Compositions, uses and methods for making them

ABSTRACT

Generally, the present invention provides novel quinolone compounds and pharmaceutical composition thereof which may inhibit cell proliferation and/or induce cell apoptosis. The present invention also provides methods of preparing such compounds and compositions, and methods of making and using the same.

CROSS-REFERENCE TO RELATED APPLICATIONS

This is a divisional application of U.S. patent application Ser. No.14/935,155, filed Nov. 6, 2015, which is a continuation-in-part ofInternational Patent Application No. PCT/US2015/030046, filed May 9,2015, which claims priority to U.S. Provisional Patent Application No.61/991,282, filed May 9, 2014; U.S. Provisional Patent Application No.62/050,202, filed Sep. 15, 2014; U.S. Provisional Patent Application No.62/054,054, filed Sep. 23, 2014; and U.S. Provisional Patent ApplicationNo. 62/128,208, filed Mar. 4, 2015, each of which are herebyincorporated by reference in their entirety.

FIELD OF THE INVENTION

The present invention provides novel compounds and pharmaceuticalcomposition thereof which may inhibit cell proliferation and/or inducecell apoptosis. The present invention also provides methods of preparingsuch compounds and compositions, and methods of making and using thesame.

BACKGROUND OF THE INVENTION

Hypertrophy of the nucleolus, the cellular site for ribosome biogenesis,has been linked to malignant transformation for more than a hundredyears. The ribosome is a RNA-protein complex that is responsible for theprotein synthesis (translation) in the cell. Carcinogenesis, with theassociated upregulation of growth and proliferation rates, requires asignificant increase in the rate of translation and hence necessitatesan increase in cellular ribosome content. Ribosome biogenesis is ahighly complex energy-consuming process in which the synthesis ofpre-ribosomal RNA by RNA Polymerase I (Pol I) serves as the ratelimiting step.

Not surprisingly, Pol I transcription in normal cells is tightlycontrolled, through the action of multiple tumor suppressor proteins(including p53, pRB and PTEN) which serve as inhibitors. The loss ofsuch control due to mutations in tumor suppressor genes or activation ofcertain oncogenic pathways, such as cMyc and PI3K/Ak/mTOR, results inthe hyperactivation of Pol I transcription that is commonly found inmalignancy.

In addition to cancer, hyperactivation of Pol I transcription has beenlinked to poor prognosis in multiple sclerosis and has been shown toplay a role in the infections cycle of certain pathologic viruses,including cytomegalovirus, hepatitis B virus and hepatitis C virus.Therefore, agents that selectively disrupt Pol I transcription areconceptually attractive as anticancer, anti-inflammatory and antiviraltherapeutics

SUMMARY OF THE INVENTION

Provided herein are novel compounds and methods of treating orpreventing any one of the diseases or conditions described hereincomprising administering a therapeutically effective amount of acompound described herein, or a pharmaceutically acceptable salt, orsolvate thereof, to a mammal in need thereof. In specific embodiments,the compound inhibits ribosome biogenesis by inhibiting POL1transcription and the disease or condition is amenable to treatment orprevention by the inhibition of POL1 transcription.

In one aspect, described herein is a method for treating or preventingcancer in a mammal comprising administering a therapeutically effectiveamount of a compound described herein, or a pharmaceutically acceptablesalt, or solvate thereof, to the mammal in need thereof. In anotheraspect, described herein is a method for treating or preventing aninflammatory disease in a mammal comprising administering atherapeutically effective amount of a compound described herein, or apharmaceutically acceptable salt, or solvate thereof, to the mammal inneed thereof.

In still another aspect, described herein is a method for treating orpreventing a proliferative disorder in a mammal comprising administeringa therapeutically effective amount of a compound described herein, or apharmaceutically acceptable salt, or solvate thereof, to the mammal inneed thereof. In another aspect, described herein is a method fortreating or preventing a disease or disorder in a mammal comprisingadministering a therapeutically effective amount of a compound describedherein, wherein the compound inhibits ribosome biogenesis by inhibitingPOL1 transcription.

Other objects, features and advantages of the compounds, methods andcompositions described herein will become apparent from the followingdetailed description. It should be understood, however, that thedetailed description and the specific examples, while indicatingspecific embodiments, are given by way of illustration only, sincevarious changes and modifications within the spirit and scope of theinstant disclosure will become apparent to those skilled in the art fromthis detailed description.

DETAILED DESCRIPTION OF THE INVENTION

Compounds

Compounds described herein, including pharmaceutically acceptable salts,prodrugs, active metabolites and pharmaceutically acceptable solvatesthereof, inhibit ribosome biogenesis by inhibiting POL1 transcription.

In one aspect, the present invention provides a compound of Formula (I),

and pharmaceutically acceptable salts, esters, prodrugs, hydrates andtautomers thereof; wherein:

-   -   Z₅ is N or CX₂;    -   Z₁, Z₂, Z₃, and Z₄ are each independently selected from N, CH,        and CR₁, provided any three N are non-adjacent; and further        provided that one or more of Z₁, Z₂, Z₃, and Z₄ is CR₁;        -   wherein each R₁ is independently an optionally substituted            C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl, C₂-C₈            heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈            acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl,            C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group, or        -   each R₁ is independently H, halo, CF₃, OR₂, NR₂R₃, NR₂OR₃,            NR₂NR₂R₃, SR₂, SOR₂, SO₂R₂, SO₂NR₂R₃, NR₂SO₂R₃, NR₂CONR₂R₃,            NR₂COOR₃, NR₂COR₃, CN, COOR₂, COOH, CONR₂R₃, OOCR₂, COR₂, or            NO₂;        -   wherein two R₁ groups on adjacent atoms may form a            carboxylic ring, heterocyclic ring, aryl or heteroaryl, each            of which may be optionally substituted and/or fused with a            cyclic ring; and        -   wherein R₂ and R₃ groups on the same atom or on adjacent            atoms can be linked to form a 3-8 membered ring, optionally            containing one or more N, O or S atoms; and each R₂ and R₃            groups, and each ring formed by linking R₂ and R₃ groups            together, is optionally substituted with one or more            substituents selected from halo, ═O, ═N—CN, ═N—OR′, ═NR′,            OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂,            NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and            NO₂,            -   wherein each R′ is independently H, C₁-C₆ alkyl, C₂-C₆                heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl,                C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂                heteroarylalkyl, each of which is optionally substituted                with one or more groups selected from halo, C₁-C₄ alkyl,                C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl,                hydroxy, amino, and ═O; wherein two R′ can be linked to                form a 3-7 membered ring optionally containing up to                three heteroatoms selected from N, O and S; or        -   each R₁ is independently —W, -L-W, —X-L-A; wherein            -   X is NR₆, O, or S;            -   W is an optionally substituted 4-7 membered azacyclic                ring, optionally containing an additional heteroatom                selected from N, O and S as a ring member;            -   L is a bond, C₁-C₁₀ alkylene, C₁-C₁₀ heteroalkylene,                C₂-C₁₀ alkenylene or C₂-C₁₀ heteroalkenylene linker,                each of which is optionally substituted with one or more                substituents selected from the group consisting of                halogen, oxo (═O), or C₁-C₆ alkyl; and            -   A is heterocycloalkyl, heteroaryl or NR₄R₅ wherein,                -   R₄ and R₅ are independently H, optionally                    substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈                    alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈                    heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀                    aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂                    heteroarylalkyl group, and R₄ and R₅ can be linked                    to form a 3-8 membered ring, optionally containing                    one or more N, O or S;                -    wherein each R₄ and R₅ groups, and each ring formed                    by linking R₄ and R₅ groups together, is optionally                    substituted with one or more substituents selected                    from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂,                    SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′,                    NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and NO₂,                    wherein each R′ is independently H, C₁-C₆ alkyl,                    C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl,                    C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or                    C₆-C₁₂ heteroarylalkyl, each of which is optionally                    substituted with one or more groups selected from                    halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl,                    C₁-C₆ heteroacyl, hydroxy, amino, and ═O; wherein                    two R′ can be linked to form a 3-7 membered ring                    optionally containing up to three heteroatoms                    selected from N, O and S;        -   wherein R₆ is H, optionally substituted C₁-C₈ alkyl, C₂-C₈            heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈            alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl,            C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂            heteroarylalkyl group,        -   wherein R₆ can be linked to R₄ or R₅ to form a 3-8 membered            ring; and        -   R₄ or R₅ is optionally substituted with one or more            substituents selected from halo, ═O, ═N—CN, ═N—OR′, ═NR′,            OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂,            NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and            NO₂, wherein each R′ is independently H, C₁-C₆ alkyl, C₂-C₆            heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl,            C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂            heteroarylalkyl, each of which is optionally substituted            with one or more groups selected from halo, C₁-C₄ alkyl,            C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy,            amino, and ═O; wherein two R′ can be linked to form a 3-7            membered ring optionally containing up to three heteroatoms            selected from N, O and S;        -   X₁ is an optionally substituted C₁-C₈ alkyl, C₂-C₈            heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈            alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl,            C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂            heteroarylalkyl group, optionally substituted with one or            more halogens, ═O, CF₃, CN, OR₇, NR₈R₉, SR₇, SO₂NR₈R₉, C₁-C₈            alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl, C₂-C₈            heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈            acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl,            C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group; or        -   X₁ is H, NR₂R₃, SOR₂, SO₂R₂, SO₂NR₂R₃, NR₂SO₂R₃, NR₂CONR₂R₃,            NR₂COOR₃, NR₂COR₃, CN, COOR₂, ester bioisostere, COOH,            carboxy bioisostere, CONR₂R₃, amide bioisostere, OOCR₂,            COR₂, or NO₂;        -   X₂ is, H, halogen, CF₃, CN, OR₇, NR₈R₉, SR₇, SO₂NR₈R₉,            C₁-C₁₀ alkyl, C₁-C₁₀ heteroalkyl, C₂-C₁₀ alkenyl, or C₂-C₁₀            heteroalkenyl, each of which is optionally substituted with            one or more halogens, ═O, or an optionally substituted 3-7            membered carbocyclic or heterocyclic ring;        -   X₃, X₄ and X₅ are each independently N or CR₁₀;            -   wherein each R₁₀ is independently an optionally                substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈                alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈                heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀                aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂                heteroarylalkyl group, or            -   each R₁ is independently H, halo, CF₃, OR₂, NR₂R₃,                NR₂OR₃, NR₂NR₂R₃, SR₂, SOR₂, SO₂R₂, SO₂NR₂R₃, NR₂SO₂R₃,                NR₂CONR₂R₃, NR₂COOR₃, NR₂COR₃, CN, COOR₂, COOH, CONR₂R₃,                OOCR₂, COR₂, or NO₂;            -   wherein R₂ and R₃ groups on the same atom or on adjacent                atoms can be linked to form a 3-8 membered ring,                optionally containing one or more N, O or S atoms; and                each R₂ and R₃ groups, and each ring formed by linking                R₂ and R₃ groups together, is optionally substituted                with one or more substituents selected from halo, ═O,                ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂,                NR′SO₂R′, NR′CONR′₂, NR′CO_(O)R′, NR′COR′, CN, COOR′,                _(C)ON(R′)₂, OOCR′, COR′, and NO₂,                -   wherein each R′ is independently H, C₁-C₆ alkyl,                    C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl,                    C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or                    C₆-C₁₂ heteroarylalkyl, each of which is optionally                    substituted with one or more groups selected from                    halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl,                    C₁-C₆ heteroacyl, hydroxy, amino, and ═O;                -   wherein two R′ can be linked to form a 3-7 membered                    ring optionally containing up to three heteroatoms                    selected from N, O and S;                -   wherein two R₁₀ groups on adjacent atoms may form a                    carboxylic ring, heterocyclic ring, aryl or                    heteroaryl, each of which is optionally substituted                    and/or fused with a cyclic ring; or            -   each R₁₀ is independently —W, -L-W, —X-L-A;                -   wherein X is NR₆, O, or S;                -   W is an optionally substituted 4-7 membered                    azacyclic ring, optionally containing an additional                    heteroatom selected from N, O and S as a ring                    member;                -   L is a bond, a C₁-C₁₀ alkylene, C₁-C₁₀                    heteroalkylene, C₂-C₁₀ alkenylene or C₂-C₁₀                    heteroalkenylene linker, each of which is optionally                    substituted with one or more substituents selected                    from the group consisting of halogen, oxo (═O), or                    C₁-C₆ alkyl; and                -   A is heterocycloalkyl, heteroaryl or NR₄R₅;                -   wherein R₄ and R₅ are independently H, optionally                    substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈                    alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈                    heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀                    aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂                    heteroarylalkyl group.

In one aspect, the present invention provides a compound of Formula1(A),

and pharmaceutically acceptable salts, esters, prodrugs, hydrates andtautomers thereof; wherein:

Y is an optionally substituted 5-6 membered carbocyclic or heterocyclicring;

X₁ is an optionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl group, optionally substituted withone or more halogens, ═O, CF₃, CN, OR₇, NR₈R₉, SR₇, SO₂NR₈R₉, C₁-C₈alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl,C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group,wherein each R₇, R₈ and R₉ is independently selected from H, C₁-C₆alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl,C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each ofwhich is optionally substituted with one or more groups selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂,NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′,COR′, and NO₂; wherein each R′ is independently H, C₁-C₆ alkyl, C₂-C₆heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of whichis optionally substituted with one or more groups selected from halo,C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy,amino, and ═O; wherein two R′ can be linked to form a 3-7 membered ringoptionally containing up to three heteroatoms selected from N, O and S;or

X₁ is H, NR₂R₃, SOR₂, SO₂R₂, SO₂NR₂R₃, NR₂SO₂R₃, NR₂CONR₂R₃, NR₂COOR₃,NR₂COR₃, CN, COOR₂, ester bioisostere, COOH, carboxy bioisostere,CONR₂R₃, amide bioisostere, OOCR₂, COR₂, or NO₂, wherein each R₂ and R₃is independently selected from H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C₁-C₄ alkyl,C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O;wherein R₂ and R₃ groups on the same atom or on adjacent atoms can belinked to form a 3-8 membered ring, optionally containing one or more N,O or S atoms; and each R₂ and R₃ groups, and each ring formed by linkingR₂ and R₃ groups together, is optionally substituted with one or moresubstituents selected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂,SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′,CON(R′)₂, OOCR′, COR′, and NO₂; wherein each R′ is independently H,C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl,each of which is optionally substituted with one or more groups selectedfrom halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl,hydroxy, amino, and ═O; wherein two R′ can be linked to form a 3-7membered ring optionally containing up to three heteroatoms selectedfrom N, O and S;

X₂ is, H, halogen, CF₃, CN, C₁-C₁₀ alkyl, C₁-C₁₀ heteroalkyl, C₂-C₁₀alkenyl, or C₂-C₁₀ heteroalkenyl, each of which is optionallysubstituted with one or more halogens, ═O, an optionally substituted 3-7membered carbocyclic or heterocyclic ring, OR₇, NR₈R₉, SR₇, or SO₂NR₈R₉;wherein each R₇, R₈ and R₉ is independently selected from H, C₁-C₆alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl,C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each ofwhich is optionally substituted with one or more groups selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂,NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′,COR′, and NO₂; wherein each R′ is independently H, C₁-C₆ alkyl, C₂-C₆heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of whichis optionally substituted with one or more groups selected from halo,C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy,amino, and ═O; wherein two R′ can be linked to form a 3-7 membered ringoptionally containing up to three heteroatoms selected from N, O and S,wherein R₇ is not hydrogen when X₂ is OR₇;

each Z₁, Z₂, Z₃, and Z₄ is independently N, CH, or CR₁, provided thatany three N atoms are non-adjacent; and further provided that one ormore of Z₁, Z₂, Z₃, and Z₄ is CR₁; wherein each R₁ is independently anoptionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl,C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl,C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, orC₆-C₁₂ heteroarylalkyl group; or each R₁ is independently H, halo, CF₃,OR₂, NR₂R₃, NR₂OR₃, NR₂NR₂R₃, SR₂, SOR₂, SO₂R₂, SO₂NR₂R₃, NR₂SO₂R₃,NR₂CONR₂R₃, NR₂COOR₃, NR₂COR₃, CN, COOR₂, COOH, CONR₂R₃, OOCR₂, COR₂, orNO₂; wherein each R₂ and R₃ is independently selected from H, C₁-C₆alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl,C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each ofwhich is optionally substituted with one or more groups selected fromhalo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl,hydroxy, amino, and ═O; wherein R₂ and R₃ groups on the same atom or onadjacent atoms can be linked to form a 3-8 membered ring, optionallycontaining one or more N, O or S atoms; and each R₂ and R₃ groups, andeach ring formed by linking R₂ and R₃ groups together, is optionallysubstituted with one or more substituents selected from halo, ═O, ═N—CN,═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂,NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and NO₂; whereineach R′ is independently H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl,C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, orC₆-C₁₂ heteroarylalkyl, each of which is optionally substituted with oneor more groups selected from halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O; wherein two R′ can belinked to form a 3-7 membered ring optionally containing up to threeheteroatoms selected from N, O and S; or

each R₁ is independently —W, -L-W, —X-L-A; wherein X is NR₆, O, or S;wherein R₆ is H, optionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl,C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl,C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl group; wherein R₆ can be linked toR₄ or R₅ to form a 3-8 membered ring and R₄ and R₅ are eachindependently optionally substituted with one or more substituentsselected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′,SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂,OOCR′, COR′, and NO₂; wherein each R′ is independently H, C₁-C₆ alkyl,C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of whichis optionally substituted with one or more groups selected from halo,C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy,amino, and ═O; wherein two R′ can be linked to form a 3-7 membered ringoptionally containing up to three heteroatoms selected from N, O and S;each W is independently an optionally substituted 4-7 membered azacyclicring, optionally containing an additional heteroatom selected from N, Oand S as a ring member; L is a bond, C₁-C₁₀ alkylene, C₁-C₁₀heteroalkylene, C₂-C₁₀ alkenylene or C₂-C₁₀ heteroalkenylene linker,each of which is optionally substituted with one or more substituentsselected from the group consisting of halogen, oxo (═O), or C₁-C₆ alkyl;and A is heterocycloalkyl, heteroaryl or NR₄R₅ wherein R₄ and R₅ areeach independently H, optionally substituted C₁-C₈ alkyl, C₂-C₈heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group; whereinR₄ and R₅ are optionally linked to form a 3-8 membered ring, optionallycontaining one or more N, O or S; and each R₄ and R₅ groups, and eachring formed by linking R₄ and R₅ groups together, is optionallysubstituted with one or more substituents selected from halo, ═O, ═N—CN,═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂,NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and NO₂; whereineach R′ is independently H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl,C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, orC₆-C₁₂ heteroarylalkyl, each of which is optionally substituted with oneor more groups selected from halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O; wherein two R′ can belinked to form a 3-7 membered ring optionally containing up to threeheteroatoms selected from N, O and S.

In some embodiments, X₁ is CN, COOR₂, or CONR₂R₃.

In some embodiments, X is NR₆.

In one aspect, the present invention provides a compound of FormulaII(A), II(B), II(C). II(D) and II (E),

or a pharmaceutically acceptable salt or ester thereof.

In some embodiments, Z₁, Z₂, Z₃ and Z₄ are independently CH or CR₁.

In some embodiments, each R₁ is independently an optionally substitutedC₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl,C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkylgroup, or each R₁ is independently halo, CF₃, OR₂, NR₂R₃, NR₂OR₃,NR₂NR₂R₃, SR₂, SOR₂, SO₂R₂, SO₂NR₂R₃, NR₂SO₂R₃, NR₂CONR₂R₃, NR₂COOR₃,NR₂COR₃, CN, COOR₂, COOH, CONR₂R₃, OOCR₂, COR₂, or NO₂.

In some embodiments, each R₁ is independently —W, -L-W, —X-L-A; whereinX is NR₆, O, or S; W is an optionally substituted 4-7 membered azacyclicring, optionally containing an additional heteroatom selected from N, Oand S as a ring member; L is a bond, C₁-C₁₀ alkylene, C₁-C₁₀heteroalkylene, C₂-C₁₀ alkenylene or C₂-C₁₀ heteroalkenylene linker,each of which is optionally substituted with one or more substituentsselected from the group consisting of halogen, oxo (═O), or C₁-C₆ alkyl;and A is heterocycloalkyl, heteroaryl, quaternary amine or NR₄R₅ whereR₄ and R₅ are independently H, optionally substituted C₁-C₈ alkyl, C₂-C₈heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group.

In some embodiments, R₄ and R₅ are linked to form a 3-8 membered ring,optionally containing one or more N, O or S; and each R₄ and R₅ groups,and each ring formed by linking R₄ and R₅ groups together, is optionallysubstituted with one or more substituents selected from halo, ═O, ═N—CN,═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂,NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and NO₂, whereineach R′ is independently H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl,C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, orC₆-C₁₂ heteroarylalkyl, each of which is optionally substituted with oneor more groups selected from halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O; wherein two R′ can belinked to form a 3-7 membered ring optionally containing up to threeheteroatoms selected from N, O and S.

In some embodiments, R₆ is H, optionally substituted C₁-C₈ alkyl, C₂-C₈heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group; or R₆ canbe linked to R₄ or R₅ to form a 3-8 membered ring; and R₄ or R₅ isoptionally substituted with one or more substituents selected from halo,═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′,NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and NO₂,wherein each R′ is independently H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl,C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C₁-C₄ alkyl,C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O;wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S.

In some embodiments, Y is an optionally substituted 5-6 memberedcarbocyclic or heterocyclic ring.

In some embodiments, X₁ is an optionally substituted C₁-C₈ alkyl, C₂-C₈heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group, or X₁ isH, NR₂R₃, SOR₂, SO₂R₂, SO₂NR₂R₃, NR₂SO₂R₃, NR₂CONR₂R₃, NR₂COOR₃,NR₂COR₃, CN, COOR₂, COOH, polar substituent, carboxy bioisostere,CONR₂R₃, OOCR₂, COR₂, or NO₂.

In some embodiments, X₂ is H, halogen, CF₃, CN, OR₇, NR₈R₉, SR₇,SO₂NR₈R₉, C₁-C₁₀ alkyl, C₁-C₁₀ heteroalkyl, C₂-C₁₀ alkenyl, or C₂-C₁₀heteroalkenyl, each of which is optionally substituted with one or morehalogens, ═O, or an optionally substituted 3-7 membered carbocyclic orheterocyclic ring, wherein R₂ and R₃ groups on the same atom or onadjacent atoms can be linked to form a 3-8 membered ring, optionallycontaining one or more N, O or S; and each R₂ and R₃ groups, and eachring formed by linking R₂ and R₃ groups together, is optionallysubstituted with one or more substituents selected from halo, ═O, ═N—CN,═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂,NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and NO₂, whereineach R′ is independently H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl,C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, orC₆-C₁₂ heteroarylalkyl, each of which is optionally substituted with oneor more groups selected from halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O; wherein two R′ can belinked to form a 3-7 membered ring optionally containing up to threeheteroatoms selected from N, O and S;

In some embodiments, the Compound of Formula (I) has the followingstructure of Formula III(A), III(B) or III(C):

or a pharmaceutically acceptable salt or ester thereof.

In some embodiments, L is a bond, C₁-C₁₀ alkylene, C₁-C₁₀heteroalkylene, C₂-C₁₀ alkenylene or C₂-C₁₀ heteroalkenylene linker,each of which is optionally substituted with one or more substituentsselected from the group consisting of halogen, oxo (═O), or C1-C6 alkyl.

In some embodiments, A is heterocycloalkyl, heteroaryl, quaternary amineor NR₄R₅ where R₄ and R₅ are independently H, optionally substitutedC₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl,C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkylgroup.

In some embodiments, R₄ and R₅ can be linked to form a 3-8 memberedring, optionally containing one or more N, O or S; and each R₄ and R₅groups, and each ring formed by linking R₄ and R₅ groups together, isoptionally substituted with one or more substituents selected from halo,═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′,NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and NO₂,wherein each R′ is independently H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl,C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C₁-C₄ alkyl,C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O;wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S.

In some embodiments, X is NR₆, O, or S.

In some embodiments, R₆ is H, optionally substituted C₁-C₈ alkyl, C₂-C₈heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group.

In some embodiments, R₆ is linked to R₄ or R₅ to form a 3-8 memberedring; and R₄ or R₅ are optionally substituted with one or moresubstituents selected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂,SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′,CON(R′)₂, OOCR′, COR′, and NO₂, wherein each R′ is independently H,C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl,each of which is optionally substituted with one or more groups selectedfrom halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl,hydroxy, amino, and ═O; wherein two R′ can be linked to form a 3-7membered ring optionally containing up to three heteroatoms selectedfrom N, O and S.

In some embodiments, X₁ is an optionally substituted C₁-C₈ alkyl, C₂-C₈heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group, or X₁ isH, NR₂R₃, SOR₂, SO₂R₂, SO₂NR₂R₃, NR₂SO₂R₃, NR₂CONR₂R₃, NR₂COOR₃,NR₂COR₃, CN, COOR₂, ester bioisostere, COOH, carboxy bioisostere,CONR₂R₃, amide bioisostere, OOCR₂, COR₂, or NO₂.

In some embodiments, X₂ is H, halogen, CF₃, CN, OR₇, NR₈R₉, SR₇,SO₂NR₈R₉, C₁-C₁₀ alkyl, C₁-C₁₀ heteroalkyl, C₂-C₁₀ alkenyl, or C₂-C₁₀heteroalkenyl, each of which is optionally substituted with one or morehalogens, ═O, or an optionally substituted 3-7 membered carbocyclic orheterocyclic ring.

In some embodiments, (U)_(n) and (U)_(m) are independently H, halogen,CF₃, CN, OR₇, NR₈R₉, SR₇, SO₂NR₈R₉, C₁-C₁₀ alkyl, C₁-C₁₀ heteroalkyl,C₂-C₁₀ alkenyl, or C₂-C₁₀ heteroalkenyl, each of which is optionallysubstituted with one or more halogens, ═O, or an optionally substituted3-7 membered carbocyclic or heterocyclic ring.

In some embodiments, R₂ and R₃ groups on the same atom or on adjacentatoms are linked to form a 3-8 membered ring, optionally containing oneor more N, O or S; and each R₂ and R₃ groups, and each ring formed bylinking R₂ and R₃ groups together, is optionally substituted with one ormore substituents selected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′,N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN,COOR′, CON(R′)₂, OOCR′, COR′, and NO₂, wherein each R′ is independentlyH, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl,each of which is optionally substituted with one or more groups selectedfrom halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl,hydroxy, amino, and ═O; wherein two R′ can be linked to form a 3-7membered ring optionally containing up to three heteroatoms selectedfrom N, O and S.

In some preferred embodiments, X₂ is H and X₁ is CONR₂R₃, wherein R₂ andR₃ are independently selected from H, C₁-C₁₀ alkyl, C₁-C₁₀ heteroalkyl,C₂-C₁₀ alkenyl, and C₂-C₁₀ heteroalkenyl, each of which is optionallysubstituted with one or more halogens, ═O, or an optionally substituted3-7 membered carbocyclic or heterocyclic ring.

In one aspect, the present invention provides a compound of FormulaIV(A), IV(B) and IV(C),

and pharmaceutically acceptable salts, esters, prodrugs, hydrates andtautomers thereof; wherein:

L is a bond, C₁-C₁₀ alkylene, C₁-C₁₀ heteroalkylene, C₂-C₁₀ alkenyleneor C₂-C₁₀ heteroalkenylene linker, each of which is optionallysubstituted with one or more substituents selected from the groupconsisting of halogen, oxo (═O), or C1-C6 alkyl;

A is heterocycloalkyl, heteroaryl or NR₄R₅ where R₄ and R₅ areindependently H, optionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl,C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl,C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl group,

R₄ and R₅ can be linked to form a 3-8 membered ring, optionallycontaining one or more N, O or S; and each R₄ and R₅ groups, and eachring formed by linking R₄ and R₅ groups together, is optionallysubstituted with one or more substituents selected from halo, ═O, ═N—CN,═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂,NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and NO₂, whereineach R′ is independently H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl,C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, orC₆-C₁₂ heteroarylalkyl, each of which is optionally substituted with oneor more groups selected from halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O; wherein two R′ can belinked to form a 3-7 membered ring optionally containing up to threeheteroatoms selected from N, O and S;

X is NR₆, O, or S;

R₆ is H, optionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl group;

R₆ can be linked to R₄ or R₅ to form a 3-8 membered ring; and R₄ or R₅is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂,NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′,COR′, and NO₂, wherein each R′ is independently H, C₁-C₆ alkyl, C₂-C₆heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of whichis optionally substituted with one or more groups selected from halo,C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy,amino, and ═O; wherein two R′ can be linked to form a 3-7 membered ringoptionally containing up to three heteroatoms selected from N, O and S;

X₁ is an optionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl group, or X₁ is H, NR₂R₃, SOR₂,SO₂R₂, SO₂NR₂R₃, NR₂SO₂R₃, NR₂CONR₂R₃, NR₂COOR₃, NR₂COR₃, CN, COOR₂,ester bioisostere, COOH, carboxy bioisostere, CONR₂R₃, amidebioisostere, OOCR₂, COR₂, or NO₂;

X₂ is H, halogen, CF₃, CN, OR₇, NR₈R₉, SR₇, SO₂NR₈R₉, C₁-C₁₀ alkyl,C₁-C₁₀ heteroalkyl, C₂-C₁₀ alkenyl, or C₂-C₁₀ heteroalkenyl, each ofwhich is optionally substituted with one or more halogens, ═O, or anoptionally substituted 3-7 membered carbocyclic or heterocyclic ring;

(U)_(n) and (U)_(m) are independently H, halogen, CF₃, CN, OR₇, NR₈R₉,SR₇, SO₂NR₈R₉, C₁-C₁₀ alkyl, C₁-C₁₀ heteroalkyl, C₂-C₁₀ alkenyl, orC₂-C₁₀ heteroalkenyl, each of which is optionally substituted with oneor more halogens, ═O, or an optionally substituted 3-7 memberedcarbocyclic or heterocyclic ring;

wherein R₂ and R₃ groups on the same atom or on adjacent atoms can belinked to form a 3-8 membered ring, optionally containing one or more N,O or S; and each R₂ and R₃ groups, and each ring formed by linking R₂and R₃ groups together, is optionally substituted with one or moresubstituents selected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂,SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′,CON(R′)₂, OOCR′, COR′, and NO₂, wherein each R′ is independently H,C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl,each of which is optionally substituted with one or more groups selectedfrom halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl,hydroxy, amino, and ═O; wherein two R′ can be linked to form a 3-7membered ring optionally containing up to three heteroatoms selectedfrom N, O and S.

In one aspect, the present invention provides a compound of FormulaV(A), V(B), V(C) and V(D),

and pharmaceutically acceptable salts, esters, prodrugs, hydrates andtautomers thereof; wherein:

L is a bond, C₁-C₁₀ alkylene, C₁-C₁₀ heteroalkylene, C₂-C₁₀ alkenyleneor C₂-C₁₀ heteroalkenylene linker, each of which is optionallysubstituted with one or more substituents selected from the groupconsisting of halogen, oxo (═O), or C1-C6 alkyl;

A is heterocycloalkyl, heteroaryl or NR₄R₅ where R₄ and R₅ areindependently H, optionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl,C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl,C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl group,

R₄ and R₅ can be linked to form a 3-8 membered ring, optionallycontaining one or more N, O or S; and each R₄ and R₅ groups, and eachring formed by linking R₄ and R₅ groups together, is optionallysubstituted with one or more substituents selected from halo, ═O, ═N—CN,═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂,NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and NO₂, whereineach R′ is independently H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl,C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, orC₆-C₁₂ heteroarylalkyl, each of which is optionally substituted with oneor more groups selected from halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O; wherein two R′ can belinked to form a 3-7 membered ring optionally containing up to threeheteroatoms selected from N, O and S;

X is NR₆, O, or S;

R₆ is H, optionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl group;

R₆ can be linked to R₄ or R₅ to form a 3-8 membered ring; and R₄ or R₅is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂,NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′,COR′, and NO₂, wherein each R′ is independently H, C₁-C₆ alkyl, C₂-C₆heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of whichis optionally substituted with one or more groups selected from halo,C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy,amino, and ═O; wherein two R′ can be linked to form a 3-7 membered ringoptionally containing up to three heteroatoms selected from N, O and S;

X₁ is an optionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl group, or X₁ is H, NR₂R₃, SOR₂,SO₂R₂, SO₂NR₂R₃, NR₂SO₂R₃, NR₂CONR₂R₃, NR₂COOR₃, NR₂COR₃, CN, COOR₂,ester bioisostere, COOH, carboxy bioisostere, CONR₂R₃, amidebioisostere, OOCR₂, COR₂, or NO₂;

(U)_(n) and (U)_(m) are independently H, halogen, CF₃, CN, OR₇, NR₈R₉,SR₇, SO₂NR₈R₉, C₁-C₁₀ alkyl, C₁-C₁₀ heteroalkyl, C₂-C₁₀ alkenyl, orC₂-C₁₀ heteroalkenyl, each of which is optionally substituted with oneor more halogens, ═O, or an optionally substituted 3-7 memberedcarbocyclic or heterocyclic ring;

wherein R₂ and R₃ groups on the same atom or on adjacent atoms can belinked to form a 3-8 membered ring, optionally containing one or more N,O or S; and each R₂ and R₃ groups, and each ring formed by linking R₂and R₃ groups together, is optionally substituted with one or moresubstituents selected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂,SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′,CON(R′)₂, OOCR′, COR′, and NO₂, wherein each R′ is independently H,C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl,each of which is optionally substituted with one or more groups selectedfrom halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl,hydroxy, amino, and ═O; wherein two R′ can be linked to form a 3-7membered ring optionally containing up to three heteroatoms selectedfrom N, O and S.

In some embodiments, the Compound of Formula (I) has the followingstructure of Formula VI:

or a pharmaceutically acceptable salt or ester thereof.

In some embodiments, L is a bond, C₁-C₁₀ alkylene, C₁-C₁₀heteroalkylene, C₂-C₁₀ alkenylene or C₂-C₁₀ heteroalkenylene linker,each of which is optionally substituted with one or more substituentsselected from the group consisting of halogen, oxo (═O), or C1-C6 alkyl.

In some embodiments, A is heterocycloalkyl, heteroaryl, quaternary amineor NR₄R₅ wherein R₄ and R₅ are independently H, optionally substitutedC₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl,C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkylgroup, or R₄ and R₅ can be linked to form a 3-8 membered ring,optionally containing one or more N, O or S; and each R₄ and R₅ groups,and each ring formed by linking R₄ and R₅ groups together, is optionallysubstituted with one or more substituents selected from halo, ═O, ═N—CN,═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂,NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and NO₂, whereineach R′ is independently H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl,C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, orC₆-C₁₂ heteroarylalkyl, each of which is optionally substituted with oneor more groups selected from halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O; wherein two R′ can belinked to form a 3-7 membered ring optionally containing up to threeheteroatoms selected from N, O and S;

In some embodiments, X is NR₆, O, or S; wherein R₆ is H, optionallysubstituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl, C₂-C₈heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl, C₂-C₈heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂heteroarylalkyl group; or R₆ can be linked to R₄ or R₅ to form a 3-8membered ring.

In some embodiments, X₁ is an optionally substituted C₁-C₈ alkyl, C₂-C₈heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group, or X₁ isH, NR₂R₃, SOR₂, SO₂R₂, SO₂NR₂R₃, NR₂SO₂R₃, NR₂CONR₂R₃, NR₂COOR₃,NR₂COR₃, CN, COOR₂, ester bioisostere, COOH, carboxy bioisostere,CONR₂R₃, amide bioisostere, OOCR₂, COR₂, or NO₂, wherein R₂ and R₃groups on the same atom or on adjacent atoms can be linked to form a 3-8membered ring, optionally containing one or more N, O or S; and each R₂and R₃ groups, and each ring formed by linking R₂ and R₃ groupstogether, is optionally substituted with one or more substituentsselected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′,SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂,OOCR′, COR′, and NO₂, wherein each R′ is independently H, C₁-C₆ alkyl,C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of whichis optionally substituted with one or more groups selected from halo,C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy,amino, and ═O; wherein two R′ can be linked to form a 3-7 membered ringoptionally containing up to three heteroatoms selected from N, O and S.

In some embodiments, X₂ is H, halogen, CF₃, CN, OR₇, NR₈R₉, SR₇,SO₂NR₈R₉, C₁-C₁₀ alkyl, C₁-C₁₀ heteroalkyl, C₂-C₁₀ alkenyl, or C₂-C₁₀heteroalkenyl, each of which is optionally substituted with one or morehalogens, ═O, or an optionally substituted 3-7 membered carbocyclic orheterocyclic ring.

In some embodiments, (U)_(n) and (U)_(m) are independently H, halogen,CF₃, CN, OR₇, NR₈R₉, SR₇, SO₂NR₈R₉, C₁-C₁₀ alkyl, C₁-C₁₀ heteroalkyl,C₂-C₁₀ alkenyl, or C₂-C₁₀ heteroalkenyl, each of which is optionallysubstituted with one or more halogens, ═O, or an optionally substituted3-7 membered carbocyclic or heterocyclic ring.

In one aspect, the present invention provides a compound of FormulaVII(A) and VII(B),

and pharmaceutically acceptable salts, esters, prodrugs, hydrates andtautomers thereof; wherein:

B₁ is a bond or C═O, B₂ is X-L-A

L is a bond, C₁-C₁₀ alkylene, C₁-C₁₀ heteroalkylene, C₂-C₁₀ alkenyleneor C₂-C₁₀ heteroalkenylene linker, each of which is optionallysubstituted with one or more substituents selected from the groupconsisting of halogen, oxo (═O), or C1-C6 alkyl;

A is heterocycloalkyl, heteroaryl or NR₄R₅ wherein R₄ and R₅ areindependently H, optionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl,C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl,C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl group, or R₄ and R₅ can be linkedto form a 3-8 membered ring, optionally containing one or more N, O orS; and each R₄ and R₅ groups, and each ring formed by linking R₄ and R₅groups together, is optionally substituted with one or more substituentsselected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′,SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂,OOCR′, COR′, and NO₂, wherein each R′ is independently H, C₁-C₆ alkyl,C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of whichis optionally substituted with one or more groups selected from halo,C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy,amino, and ═O; wherein two R′ can be linked to form a 3-7 membered ringoptionally containing up to three heteroatoms selected from N, O and S;

X is CR₆R₆, NR₆, O, or S; wherein R₆ is H, optionally substituted C₁-C₈alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl,C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group; orR₆ can be linked to R₄ or R₅ to form a 3-8 membered ring;

X₂ is H, halogen, CF₃, CN, OR₇, NR₈R₉, SR₇, SO₂NR₈R₉, C₁-C₁₀ alkyl,C₁-C₁₀ heteroalkyl, C₂-C₁₀ alkenyl, or C₂-C₁₀ heteroalkenyl, each ofwhich is optionally substituted with one or more halogens, ═O, or anoptionally substituted 3-7 membered carbocyclic or heterocyclic ring.

(U)_(n) and (U)_(m) are independently H, halogen, CF₃, CN, OR₇, NR₈R₉,SR₇, SO₂NR₈R₉, C₁-C₁₀ alkyl, C₁-C₁₀ heteroalkyl, C₂-C₁₀ alkenyl, orC₂-C₁₀ heteroalkenyl, each of which is optionally substituted with oneor more halogens, ═O, or an optionally substituted 3-7 memberedcarbocyclic or heterocyclic ring.

In one aspect, the present invention provides a compound of FormulaVIII(A), VIII(B) and VIII(C),

and pharmaceutically acceptable salts, esters, prodrugs, hydrates andtautomers thereof; wherein:

B₁ is a bond or C═O, B₂ is X-L-A

L is a bond, C₁-C₁₀ alkylene, C₁-C₁₀ heteroalkylene, C₂-C₁₀ alkenyleneor C₂-C₁₀ heteroalkenylene linker, each of which is optionallysubstituted with one or more substituents selected from the groupconsisting of halogen, oxo (═O), or C1-C6 alkyl;

A is heterocycloalkyl, heteroaryl or NR₄R₅ wherein R₄ and R₅ areindependently H, optionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl,C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl,C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl group, or R₄ and R₅ can be linkedto form a 3-8 membered ring, optionally containing one or more N, O orS; and each R₄ and R₅ groups, and each ring formed by linking R₄ and R₅groups together, is optionally substituted with one or more substituentsselected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′,SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂,OOCR′, COR′, and NO₂, wherein each R′ is independently H, C₁-C₆ alkyl,C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of whichis optionally substituted with one or more groups selected from halo,C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy,amino, and ═O; wherein two R′ can be linked to form a 3-7 membered ringoptionally containing up to three heteroatoms selected from N, O and S;

X is CR₆R₆, NR₆, O, or S; wherein R₆ is H, optionally substituted C₁-C₈alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl,C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group; orR₆ can be linked to R₄ or R₅ to form a 3-8 membered ring;

X₂ is H, halogen, CF₃, CN, OR₇, NR₈R₉, SR₇, SO₂NR₈R₉, C₁-C₁₀ alkyl,C₁-C₁₀ heteroalkyl, C₂-C₁₀ alkenyl, or C₂-C₁₀ heteroalkenyl, each ofwhich is optionally substituted with one or more halogens, ═O, or anoptionally substituted 3-7 membered carbocyclic or heterocyclic ring.

(U)_(n) and (U)_(m) are independently H, halogen, CF₃, CN, OR₇, NR₈R₉,SR₇, SO₂NR₈R₉, C₁-C₁₀ alkyl, C₁-C₁₀ heteroalkyl, C₂-C₁₀ alkenyl, orC₂-C₁₀ heteroalkenyl, each of which is optionally substituted with oneor more halogens, ═O, or an optionally substituted 3-7 memberedcarbocyclic or heterocyclic ring.

Any combination of the groups described above for the various variablesis contemplated herein. Throughout the specification, groups andsubstituents thereof are chosen by one skilled in the field to providestable moieties and compounds.

In one aspect, described herein are pharmaceutical compositionscomprising a compound described herein, or a pharmaceutically acceptablesalt, or solvate thereof, and at least one pharmaceutically acceptableexcipient. In some embodiments, the pharmaceutical composition isformulated for administration to a mammal by intravenous administration,subcutaneous administration, oral administration, inhalation, nasaladministration, dermal administration, or ophthalmic administration. Insome embodiments, the pharmaceutical composition is in the form of atablet, a pill, a capsule, a liquid, a suspension, a gel, a dispersion,a solution, an emulsion, an ointment, or a lotion.

In one aspect, described herein is a method of treating or preventingany one of the diseases or conditions described herein comprisingadministering a therapeutically effective amount of a compound describedherein, or a pharmaceutically acceptable salt, or solvate thereof, to amammal in need thereof.

In another aspect, described herein is a method for treating orpreventing cancer, or fibrosis, or combinations thereof in a mammalcomprising administering a therapeutically effective amount of acompound described herein, or a pharmaceutically acceptable salt, orsolvate thereof, to the mammal in need thereof.

In one aspect, described herein is a method for treating or preventingcancer in a mammal comprising administering a therapeutically effectiveamount of a compound described herein, or a pharmaceutically acceptablesalt, or solvate thereof, to the mammal in need thereof. In someembodiments, the cancer is amenable to treatment with an inhibitor ofPOL1 transcription. In some embodiments, the method further comprisesadministering a second therapeutic agent to the mammal in addition tothe compound described herein, or a pharmaceutically acceptable salt, orsolvate thereof.

In one aspect, described herein is a method for treating or preventingan inflammatory disease in a mammal comprising administering atherapeutically effective amount of a compound described herein, or apharmaceutically acceptable salt, or solvate thereof, to the mammal inneed thereof. In some embodiments, the inflammatory disease is amenableto treatment with an inhibitor of POL1 transcription. In someembodiments, the method further comprises administering a secondtherapeutic agent to the mammal in addition to the compound describedherein, or a pharmaceutically acceptable salt, or solvate thereof.

In one aspect, described herein is a method for treating or preventing aproliferative disorder in a mammal comprising administering atherapeutically effective amount of a compound described herein, or apharmaceutically acceptable salt, or solvate thereof, to the mammal inneed thereof. In some embodiments, the proliferative disorder isamenable to treatment with an inhibitor of POL1 transcription. In someembodiments, the method further comprises administering a secondtherapeutic agent to the mammal in addition to the compound describedherein, or a pharmaceutically acceptable salt, or solvate thereof.

In one aspect, described herein is a method for treating or preventing adisease or disorder in a mammal comprising administering atherapeutically effective amount of a compound described herein, whereinthe compound inhibits ribosome biogenesis by inhibiting POL1transcription. In some embodiments, the method further comprisesadministering a second therapeutic agent to the mammal in addition tothe compound described herein, or a pharmaceutically acceptable salt, orsolvate thereof.

In any of the aforementioned aspects are further embodiments in whichthe effective amount of the compound described herein, or apharmaceutically acceptable salt thereof, is: (a) systemicallyadministered to the mammal; and/or (b) administered orally to themammal; and/or (c) intravenously administered to the mammal; and/or (d)administered by inhalation; and/or (e) t administered by nasaladministration; or and/or (f) administered by injection to the mammal;and/or (g) administered topically to the mammal; and/or (h) administeredby ophthalmic administration; and/or (i) administered rectally to themammal; and/or (j) administered non-systemically or locally to themammal.

In any of the aforementioned aspects are further embodiments comprisingsingle administrations of the effective amount of the compound,including further embodiments in which the compound is administered oncea day to the mammal or the compound is administered to the mammalmultiple times over the span of one day. In some embodiments, thecompound is administered on a continuous dosing schedule. In someembodiments, the compound is administered on a continuous daily dosingschedule.

In any of the aforementioned aspects involving the treatment of POL1transcription related diseases or conditions are further embodimentscomprising administering at least one additional agent in addition tothe administration of a compound described herein, or a pharmaceuticallyacceptable salt thereof. In various embodiments, each agent isadministered in any order, including simultaneously.

In any of the embodiments disclosed herein, the mammal is a human.

In some embodiments, compounds provided herein are administered to ahuman.

In some embodiments, compounds provided herein are orally administered.

Articles of manufacture, which include packaging material, a compounddescribed herein, or a pharmaceutically acceptable salt thereof, withinthe packaging material, and a label that indicates that the compound orcomposition, or pharmaceutically acceptable salt, tautomers,pharmaceutically acceptable N-oxide, pharmaceutically active metabolite,pharmaceutically acceptable prodrug, or pharmaceutically acceptablesolvate thereof, is used for inhibiting ribosome biogenesis byinhibiting POL1 transcription, or for the treatment, prevention oramelioration of one or more symptoms of a disease or condition thatwould benefit from inhibition of POL1 transcription, are provided.

In one aspect, compounds described herein are in the form ofpharmaceutically acceptable salts. As well, active metabolites of thesecompounds having the same type of activity are included in the scope ofthe present disclosure. In addition, the compounds described herein canexist in unsolvated as well as solvated forms with pharmaceuticallyacceptable solvents such as water, ethanol, and the like. The solvatedforms of the compounds presented herein are also considered to bedisclosed herein.

“Pharmaceutically acceptable,” as used herein, refers a material, suchas a carrier or diluent, which does not abrogate the biological activityor properties of the compound, and is relatively nontoxic, i.e., thematerial is administered to an individual without causing undesirablebiological effects or interacting in a deleterious manner with any ofthe components of the composition in which it is contained.

The term “pharmaceutically acceptable salt” refers to a form of atherapeutically active agent that consists of a cationic form of thetherapeutically active agent in combination with a suitable anion, or inalternative embodiments, an anionic form of the therapeutically activeagent in combination with a suitable cation. Handbook of PharmaceuticalSalts: Properties, Selection and Use. International Union of Pure andApplied Chemistry, Wiley-VCH 2002. S. M. Berge, L. D. Bighley, D. C.Monkhouse, J. Pharm. Sci. 1977, 66, 1-19. P. H. Stahl and C. G. Wermuth,editors, Handbook of Pharmaceutical Salts: Properties, Selection andUse, Weinheim/Zürich:Wiley-VCH/VHCA, 2002. Pharmaceutical saltstypically are more soluble and more rapidly soluble in stomach andintestinal juices than non-ionic species and so are useful in soliddosage forms. Furthermore, because their solubility often is a functionof pH, selective dissolution in one or another part of the digestivetract is possible and this capability can be manipulated as one aspectof delayed and sustained release behaviors. Also, because thesalt-forming molecule can be in equilibrium with a neutral form, passagethrough biological membranes can be adjusted.

In some embodiments, pharmaceutically acceptable salts are obtained byreacting a compound described herein with an acid. In some embodiments,the compound described herein (i.e. free base form) is basic and isreacted with an organic acid or an inorganic acid. Inorganic acidsinclude, but are not limited to, hydrochloric acid, hydrobromic acid,sulfuric acid, phosphoric acid, nitric acid, and metaphosphoric acid.Organic acids include, but are not limited to, 1-hydroxy-2-naphthoicacid; 2,2-dichloroacetic acid; 2-hydroxyethanesulfonic acid;2-oxoglutaric acid; 4-acetamidobenzoic acid; 4-aminosalicylic acid;acetic acid; adipic acid; ascorbic acid (L); aspartic acid (L);benzenesulfonic acid; benzoic acid; camphoric acid (+);camphor-10-sulfonic acid (+); capric acid (decanoic acid); caproic acid(hexanoic acid); caprylic acid (octanoic acid); carbonic acid; cinnamicacid; citric acid; cyclamic acid; dodecylsulfuric acid;ethane-1,2-disulfonic acid; ethanesulfonic acid; formic acid; fumaricacid; galactaric acid; gentisic acid; glucoheptonic acid (D); gluconicacid (D); glucuronic acid (D); glutamic acid; glutaric acid;glycerophosphoric acid; glycolic acid; hippuric acid; isobutyric acid;lactic acid (DL); lactobionic acid; lauric acid; maleic acid; malic acid(−L); malonic acid; mandelic acid (DL); methanesulfonic acid;naphthalene-1,5-disulfonic acid; naphthalene-2-sulfonic acid; nicotinicacid; oleic acid; oxalic acid; palmitic acid; pamoic acid; phosphoricacid; proprionic acid; pyroglutamic acid (−L); salicylic acid; sebacicacid; stearic acid; succinic acid; sulfuric acid; tartaric acid (+L);thiocyanic acid; toluenesulfonic acid (p); and undecylenic acid.

In some embodiments, a compound described herein is prepared as achloride salt, sulfate salt, bromide salt, mesylate salt, maleate salt,citrate salt or phosphate salt. In some embodiments, a compounddescribed herein is prepared as a hydrochloride salt.

In some embodiments, pharmaceutically acceptable salts are obtained byreacting a compound described herein with a base. In some embodiments,the compound described herein is acidic and is reacted with a base. Insuch situations, an acidic proton of the compound described herein isreplaced by a metal ion, e.g., lithium, sodium, potassium, magnesium,calcium, or an aluminum ion. In some cases, compounds described hereincoordinate with an organic base, such as, but not limited to,ethanolamine, diethanolamine, triethanolamine, tromethamine, meglumine,N-methylglucamine, dicyclohexylamine, tris(hydroxymethyl)methylamine. Inother cases, compounds described herein form salts with amino acids suchas, but not limited to, arginine, lysine, and the like. Acceptableinorganic bases used to form salts with compounds that include an acidicproton, include, but are not limited to, aluminum hydroxide, calciumhydroxide, potassium hydroxide, sodium carbonate, potassium carbonate,sodium hydroxide, lithium hydroxide, and the like. In some embodiments,the compounds provided herein are prepared as a sodium salt, calciumsalt, potassium salt, magnesium salt, meglumine salt, N-methylglucaminesalt or ammonium salt. In some embodiments, the compounds providedherein are prepared as a sodium salt.

It should be understood that a reference to a pharmaceuticallyacceptable salt includes the solvent addition forms. In someembodiments, solvates contain either stoichiometric ornon-stoichiometric amounts of a solvent, and are formed during theprocess of crystallization with pharmaceutically acceptable solventssuch as water, ethanol, and the like. Hydrates are formed when thesolvent is water, or alcoholates are formed when the solvent is alcohol.Solvates of compounds described herein are conveniently prepared orformed during the processes described herein. In addition, the compoundsprovided herein optionally exist in unsolvated as well as solvatedforms.

The methods and formulations described herein include the use ofN-oxides (if appropriate), crystalline forms (also known as polymorphs),or pharmaceutically acceptable salts of compounds described herein, aswell as active metabolites of these compounds having the same type ofactivity.

In some embodiments, sites on the organic radicals (e.g. alkyl groups,aromatic rings) of compounds described herein are susceptible to variousmetabolic reactions. Incorporation of appropriate substituents on theorganic radicals will reduce, minimize or eliminate this metabolicpathway. In specific embodiments, the appropriate substituent todecrease or eliminate the susceptibility of the aromatic ring tometabolic reactions is, by way of example only, a halogen, deuterium, analkyl group, a haloalkyl group, or a deuteroalkyl group.

In another embodiment, the compounds described herein are labeledisotopically (e.g. with a radioisotope) or by another other means,including, but not limited to, the use of chromophores or fluorescentmoieties, bioluminescent labels, or chemiluminescent labels.

Compounds described herein include isotopically-labeled compounds, whichare identical to those recited in the various formulae and structurespresented herein, but for the fact that one or more atoms are replacedby an atom having an atomic mass or mass number different from theatomic mass or mass number usually found in nature. Examples of isotopesthat can be incorporated into the present compounds include isotopes ofhydrogen, carbon, nitrogen, oxygen, fluorine and chlorine, such as, forexample, ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³⁵S, ¹⁸F, ³⁶Cl. In one aspect,isotopically-labeled compounds described herein, for example those intowhich radioactive isotopes such as ³H and ¹⁴C are incorporated, areuseful in drug and/or substrate tissue distribution assays. In oneaspect, substitution with isotopes such as deuterium affords certaintherapeutic advantages resulting from greater metabolic stability, suchas, for example, increased in vivo half-life or reduced dosagerequirements.

In some embodiments, the compounds described herein possess one or morestereocenters and each stereocenter exists independently in either the Ror S configuration. The compounds presented herein include alldiastereomeric, enantiomeric, atropisomers, and epimeric forms as wellas the appropriate mixtures thereof. The compounds and methods providedherein include all cis, trans, syn, anti, entgegen (E), and zusammen (Z)isomers as well as the appropriate mixtures thereof.

Individual stereoisomers are obtained, if desired, by methods such as,stereoselective synthesis and/or the separation of stereoisomers bychiral chromatographic columns. In certain embodiments, compoundsdescribed herein are prepared as their individual stereoisomers byreacting a racemic mixture of the compound with an optically activeresolving agent to form a pair of diastereoisomeric compounds/salts,separating the diastereomers and recovering the optically pureenantiomers. In some embodiments, resolution of enantiomers is carriedout using covalent diastereomeric derivatives of the compounds describedherein. In another embodiment, diastereomers are separated byseparation/resolution techniques based upon differences in solubility.In other embodiments, separation of steroisomers is performed bychromatography or by the forming diastereomeric salts and separation byrecrystallization, or chromatography, or any combination thereof. JeanJacques, Andre Collet, Samuel H. Wilen, “Enantiomers, Racemates andResolutions”, John Wiley And Sons, Inc., 1981. In some embodiments,stereoisomers are obtained by stereoselective synthesis.

In some embodiments, compounds described herein are prepared asprodrugs. A “prodrug” refers to an agent that is converted into theparent drug in vivo. Prodrugs are often useful because, in somesituations, they are easier to administer than the parent drug. Theyare, for instance, bioavailable by oral administration whereas theparent is not. Further or alternatively, the prodrug also has improvedsolubility in pharmaceutical compositions over the parent drug. In someembodiments, the design of a prodrug increases the effective watersolubility. An example, without limitation, of a prodrug is a compounddescribed herein, which is administered as an ester (the “prodrug”) butthen is metabolically hydrolyzed to provide the active entity. A furtherexample of a prodrug is a short peptide (polyaminoacid) bonded to anacid group where the peptide is metabolized to reveal the active moiety.In certain embodiments, upon in vivo administration, a prodrug ischemically converted to the biologically, pharmaceutically ortherapeutically active form of the compound. In certain embodiments, aprodrug is enzymatically metabolized by one or more steps or processesto the biologically, pharmaceutically or therapeutically active form ofthe compound.

Prodrugs of the compounds described herein include, but are not limitedto, esters, ethers, carbonates, thiocarbonates, N-acyl derivatives,N-acyloxyalkyl derivatives, quaternary derivatives of tertiary amines,N-Mannich bases, Schiff bases, amino acid conjugates, phosphate esters,and sulfonate esters. For example, see Design of Prodrugs, Bundgaard, A.Ed., Elseview, 1985 and Method in Enzymology, Widder, K. et al., Ed.;Academic, 1985, vol. 42, p. 309-396; Bundgaard, H. “Design andApplication of Prodrugs” in A Textbook of Drug Design and Development,Krosgaard-Larsen and H. Bundgaard, Ed., 1991, Chapter 5, p. 113-191; andBundgaard, H., Advanced Drug Delivery Review, 1992, 8, 1-38, each ofwhich is incorporated herein by reference.

In some embodiments, a hydroxyl group in the compounds disclosed hereinis used to form a prodrug, wherein the hydroxyl group is incorporatedinto an acyloxyalkyl ester, alkoxycarbonyloxyalkyl ester, alkyl ester,aryl ester, phosphate ester, sugar ester, ether, and the like. In someembodiments, a hydroxyl group in the compounds disclosed herein is aprodrug wherein the hydroxyl is then metabolized in vivo to provide acarboxylic acid group. In some embodiments, a carboxyl group is used toprovide an ester or amide (i.e. the prodrug), which is then metabolizedin vivo to provide a carboxylic acid group. In some embodiments,compounds described herein are prepared as alkyl ester prodrugs.

Prodrug forms of the herein described compounds, wherein the prodrug ismetabolized in vivo to produce a compound described herein as set forthherein are included within the scope of the claims. In some cases, someof the herein-described compounds are a prodrug for another derivativeor active compound.

In additional or further embodiments, the compounds described herein aremetabolized upon administration to an organism in need to produce ametabolite that is then used to produce a desired effect, including adesired therapeutic effect.

A “metabolite” of a compound disclosed herein is a derivative of thatcompound that is formed when the compound is metabolized. The term“active metabolite” refers to a biologically active derivative of acompound that is formed when the compound is metabolized. The term“metabolized,” as used herein, refers to the sum of the processes(including, but not limited to, hydrolysis reactions and reactionscatalyzed by enzymes) by which a particular substance is changed by anorganism. Thus, enzymes may produce specific structural alterations to acompound. For example, cytochrome P450 catalyzes a variety of oxidativeand reductive reactions while uridine diphosphate glucuronyltransferasescatalyze the transfer of an activated glucuronic-acid molecule toaromatic alcohols, aliphatic alcohols, carboxylic acids, amines and freesulphydryl groups. Metabolites of the compounds disclosed herein areoptionally identified either by administration of compounds to a hostand analysis of tissue samples from the host, or by incubation ofcompounds with hepatic cells in vitro and analysis of the resultingcompounds.

Synthesis of Compounds

Compounds described herein are synthesized using standard synthetictechniques or using methods know in the art in combination with methoddescribed herein.

General synthetic method for preparing intermediates and compoundsdescribed herein is shown in Scheme 1.

Compounds of formula C are formed by reacting compounds of formula Awith compound of formula B under known condensation conditions (See,e.g., Eur. J. Org. Chem., 2004, 546-551, J. Org. Chem., 2006, 71,5440-5447, Synthesis, 2003, 555-559, Eur. J. Org. Chem., 2006,3767-3770, Org. Lett., 2013, 15, 1854-1857, J. Org. Chem., 2007, 72,9854-9856, Synlett, 2011, 1723-1726, Org. Lett., 2013, 15, 4564-4567,Eur. J. Org. Chem., 2006, 3767-3770).

In certain instances the reaction of compounds A and compounds B lead inone step to compounds C. In other instances two steps are needed to formcompounds C from compounds A and B. First step is the formation of thecondensation products followed by nucleophilic reaction underappropriate conditions.

Another general synthetic method for preparing starting materialsdescribed herein is shown in Exemplary Scheme 2.

Exemplary starting materials useful in Exemplary Scheme 2 include:

As used herein, “EWG” refers to an electron withdrawing group. Asunderstood in the art, an electron withdrawing group is an atom or groupthat draws electron density from neighboring atoms towards itself,usually by resonance or inductive effects.

In some embodiments, the preparation of the compounds is made with thesequence of steps shown in Exemplary Scheme 3.

Compound 3 is prepared from the reaction of reagents 1 and 2 usingknoevenagel condensation. Compound 4 is prepared by reacting Compound 3with reagent A-L-XH. The formation Compound 5 from Compound 4 is knownin the art. Compound 6 is prepared by the coupling reaction of acid 5and amines.

In some embodiments, the preparation of the compounds is made with thesequence of steps shown in Exemplary Scheme 4.

In some embodiments, the preparation of the compounds is made with thesequence of steps shown in Exemplary Scheme 5.

In some embodiments, the preparation of the compounds is made with thesequence of steps shown in Exemplary Scheme 6.

In some embodiments, the preparation of the compounds is made with thesequence of steps shown in Exemplary Scheme 7.

In some embodiments, the preparation of the compounds is made with thesequence of steps shown in Exemplary Scheme 8.

The aldehyde 18 (prepared, for example, according to U.S. Pat. No.8,637,529 or 7,998,978) is reacted with2,6-dichloropyridine-3-carboxyaldehyde and N-methylhomopiperazine inethanol to give compound 19. Alternatively, the reaction of aldehyde 18with ethyl benzimidazole-2-acetate and N-methylhomopiperazine in ethanolgives compound 20.

The ester group in compound 20 can be hydrolyzed under acidic or basicconditions to form the acid analog which can be coupled to an amine, forexample methyl amine, under normal amide coupling to give compound 21.

In some embodiments, the preparation of the compounds is made with thesequence of steps shown in Exemplary Scheme 9.

The aldehyde 22 (prepared according to U.S. Pat. No. 8,518,952) isreacted with 2,6-dichloropyridine-3-carboxyaldehyde andN-methylhomopiperazine in ethanol to give compound 23. Alternatively,the reaction of aldehyde 22 with ethyl benzimidazole-2-acetate andN-methylhomopiperazine in ethanol gives compound 24.

The ester group in compound 20 can be hydrolyzed under acidic or basicconditions to form the acid analog which can be coupled to an amine, forexample methyl amine, under normal amide coupling conditions to givecompound 25.

In some embodiments, the preparation of the compounds is made with thesequence of steps shown in Exemplary Scheme 10.

Non-limiting specific examples of A-L-XH in the compounds describedherein are illustrated in Figure 1.

Non-limiting specific examples of R₃R₂NH in the compounds describedherein are illustrated in Figure 2.

Non-limiting specific examples of compounds of Formula II(C) asdescribed herein are illustrated in Figure 3.

Non-limiting specific examples of compounds of Formula II(D) asdescribed herein are illustrated in Figure 4.

Non-limiting specific examples of compounds described herein areillustrated in Figure 5.

A method for preparing compounds of formula VI(B) are illustrated inExemplary Scheme 11

Method for preparing compounds of Formula VI(A) are illustrated inExemplary Scheme 12.

Compounds 30 and 34 can be prepared as described in exemplary scheme 8.The methyl ester group in compound 28 and 34 can be converted to ahydroxyl using a reducing agent such as LiBH₄ under appropriateconditions. The transformation of hydroxyl group in compounds 31 and 37to a leaving group, followed by nucleophilic substitution leads tocompounds 33 and 39 respectively.

Figure 6 provides non-limiting representative examples of substitutedchloropyridinecarboxaldehyde.

Method for preparing compounds of formula VI(A) are illustrated inExemplary Scheme 13.

Compound 35 reacts with N,O-dimethylhydroxylamine hydrochloride underappropriate amide coupling conditions to give compound 40 and subsequenttreatment of this compound with an organometallic reagent such as MeMgBrleads to compound 41. The treatment of compound 41 with brominatingreagent such as N-Bromosuccinimide gives compound 42. The nucleophilicsubstitution with amines leads to compound 43.

A method for preparing compounds of formula V(A), V(B), V(C) and V(D)are illustrated in Exemplary Scheme 14.

Compound 44 is prepared according to U.S. Pat. No. 7,816,524. Thechlorination of compound 44 using chlorinating agent such as POCl₃ leadsto compound 45. Compound 45 undergoes nucleophilic substitution withHX-L-A (as defined above) to yield 46.

Compounds described herein are also prepared according to ExemplaryScheme 15.

Compound 47 can be prepared as described in exemplary scheme 14.Compound 47 reacts with N,N-Dimethylformamide dimethyl acetal to givecompound 48. Compound 48 reacts with substituted hydrazine orsubstituted amidine to yield compound 49 and 50 respectively.

Figure 7 provides non-limiting representative examples of R₂R₁NH used inexemplary schemes 12, 13, 14, and 15.

Compounds described herein are also prepared according to ExemplaryScheme 16.

Compound 3 is prepared from the reaction of reagents 1 and 2 inappropriate solvent and appropriate temperature in the presence of anamine. The acid 4 can be prepared from hydrolysis of compound 3(X=COOEt) and subsequent amide coupling leads to compound 5.

Non-limiting specific examples that can be prepared by the method setforth in Exemplary Scheme 16 include:

Other specific non-limiting examples prepared by the methods describedherein include:

-   4-(4-Methyl-piperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-Methyl-4-(4-methyl-piperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-(4-Methyl-piperazin-1-yl)-2-trifluoromethyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid hydroxyamide-   2-Methyl-4-(4-methyl-piperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid hydroxyamide-   4-(4-Methyl-piperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid hydroxyamide-   4-(4-Methyl-piperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methoxy-amide-   2-Methyl-4-(4-methyl-piperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methoxy-amide-   4-(4-Methyl-piperazin-1-yl)-2-trifluoromethyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methoxy-amide-   4-(3-Amino-piperidin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-(3-Amino-piperidin-1-yl)-2-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-(3-Amino-piperidin-1-yl)-2-trifluoromethyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-[2-(3-Fluoro-pyrrolidin-1-yl)-ethylamino]-2-trifluoromethyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-[2-(3-Fluoro-pyrrolidin-1-yl)-ethylamino]-2-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-[2-(3-Fluoro-pyrrolidin-1-yl)-ethylamino]-2-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile-   2-Methyl-4-(4-methyl-piperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile-   2-Methyl-4-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile-   4-(4-Ethyl-[1,4]diazepan-1-yl)-2-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile-   4-(4-Isopropyl-[1,4]diazepan-1-yl)-2-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile-   4-[1,4]Diazepan-1-yl-2-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile-   4-[1,4]Diazepan-1-yl-2-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-[1,4]Diazepan-1-yl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   [4-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluoren-6-yl]-morpholin-4-yl-methanone-   4-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-hydroxy-ethyl)-amide-   4-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-methoxy-ethyl)-amide-   4-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-methoxy-1-methyl-ethyl)-amide-   2-Methyl-4-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-methoxy-1-methyl-ethyl)-amide-   2-Methyl-4-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-methoxy-ethyl)-amide-   4-(3,5-Dimethyl-piperazin-1-yl)-2-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-methoxy-ethyl)-amide-   4-(3,5-Dimethyl-piperazin-1-yl)-2-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid oxetan-3-ylamide-   4-(3,5-Dimethyl-piperazin-1-yl)-2-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (pyridin-3-ylmethyl)-amide-   10-Chloro-4-(3,5-dimethyl-piperazin-1-yl)-2-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   9-Chloro-4-(3,5-dimethyl-piperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-(3,5-Dimethyl-piperazin-1-yl)-9,10-dimethyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-(2-Morpholin-4-yl-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-(2-Diethylamino-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-[(1-Methyl-1H-imidazol-4-ylmethyl)-amino]-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-(2-Imidazol-1-yl-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-[4-(2-Methoxy-ethyl)-piperazin-1-yl]-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-[4-(2-Methoxy-ethyl)-piperazin-1-yl]-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid isopropylamide-   4-(4-Methyl-[1,4]diazepan-1-yl)-6-[1,3,4]oxadiazol-2-yl-1,7,11b-triaza-benzo[c]fluorene-   4-(4-Methyl-[1,4]diazepan-1-yl)-6-(5-methyl-[1,3,4]oxadiazol-2-yl)-1,7,11b-triaza-benzo[c]fluorene-   4-(4-Methyl-[1,4]diazepan-1-yl)-6-(4H-[1,2,4]triazol-3-yl)-1,7,11b-triaza-benzo[c]fluorine-   4-(4-Methyl-[1,4]diazepan-1-yl)-6-(1H-tetrazol-5-yl)-1,7,11b-triaza-benzo[c]fluorine-   4-(4-Methyl-[1,4]diazepan-1-yl)-6-oxazol-2-yl-1,7,11b-triaza-benzo[c]fluorine-   4-(3,5-Dimethyl-piperazin-1-yl)-6-oxazol-2-yl-1,7,11b-triaza-benzo[c]fluorine-   4-(3,5-Dimethyl-piperazin-1-yl)-6-[1,3,4]oxadiazol-2-yl-1,7,11b-triaza-benzo[c]fluorine-   4-(3,5-Dimethyl-piperazin-1-yl)-6-(4H-[1,2,4]triazol-3-yl)-1,7,11b-triaza-benzo[c]fluorine-   4-(3,5-Dimethyl-piperazin-1-yl)-6-(5-methyl-4H-[1,2,4]triazol-3-yl)-1,7,11b-triaza-benzo[c]fluorene-   3-Fluoro-2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   3-Fluoro-2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid isopropylamide-   2-(3,5-Dimethyl-piperazin-1-yl)-3-fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   3-Fluoro-2-(4-methyl-piperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   3-Fluoro-2-[4-(2-methoxy-ethyl)-piperazin-1-yl]-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   3-Fluoro-2-(2-pyrrolidin-1-yl-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-(3,5-Dimethyl-piperazin-1-yl)-3-fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-(3,5-Dimethyl-piperazin-1-yl)-3-fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid isopropylamide-   2-(3,5-Dimethyl-piperazin-1-yl)-3-fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid cyclopropylamide-   2-(3,5-Dimethyl-piperazin-1-yl)-3-fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-methoxy-ethyl)-amide-   2-(3,5-Dimethyl-piperazin-1-yl)-3-fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid ethylamide-   2-(3,5-Dimethyl-piperazin-1-yl)-4-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid ethylamide-   2-(3,5-Dimethyl-piperazin-1-yl)-4-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-(3,5-Dimethyl-piperazin-1-yl)-4-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid cyclopropylamide-   2-(3,5-Dimethyl-piperazin-1-yl)-4-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-methoxy-ethyl)-amide-   2-(3,5-Dimethyl-piperazin-1-yl)-4-trifluoromethyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-methoxy-ethyl)-amide-   2-(3,5-Dimethyl-piperazin-1-yl)-4-trifluoromethyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-[1,4]Diazepan-1-yl-4-trifluoromethyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-[1,4]Diazepan-1-yl-4-methyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-[1,4]Diazepan-1-yl-3-fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-[1,4]Diazepan-1-yl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-(4-Ethyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-(4-Isopropyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid ethylamide-   2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (pyridin-3-ylmethyl)-amide-   2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-imidazol-1-yl-ethyl)-amide-   10-Chloro-2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   10-Cyano-2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   9,10-Dimethyl-2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   4-Methyl-2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-(4-Methyl-[1,4]diazepan-1-yl)-4-trifluoromethyl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   3-Fluoro-2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid hydroxyamide-   3-Fluoro-2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methoxy-amide-   3-Fluoro-2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid N′-methyl-hydrazide-   3-Fluoro-2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid amide-   2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid amide-   2-[1,4]Diazepan-1-yl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid amide-   2-[1,4]Diazepan-1-yl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid-   2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-hydroxy-1-methyl-ethyl)-amide-   2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-methoxy-1-methyl-ethyl)-amide-   [2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluoren-6-yl]-pyrrolidin-1-yl-methanone-   [2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluoren-6-yl]-(2-methyl-pyrrolidin-1-yl)-methanone-   (3-Hydroxymethyl-pyrrolidin-1-yl)-[2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluoren-6-yl]-methanone-   (3,3-Difluoro-pyrrolidin-1-yl)-[2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluoren-6-yl]-methanone-   (3-Dimethylamino-pyrrolidin-1-yl)-[2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluoren-6-yl]-methanone-   2-(3-Dimethylamino-pyrrolidin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid methylamide-   2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluoren-6-yl]-(4-methyl-piperazin-1-yl)-methanone-   2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (1-methyl-azetidin-3-yl)-amide-   2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2-pyrrolidin-1-yl-ethyl)-amide-   2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic    acid (2,2-difluoro-ethyl)-amide-   [1,4]Diazepan-1-yl-(3,7,11b-triaza-benzo[c]fluoren-6-yl)-methanone-   (4-Methyl-[1,4]diazepan-1-yl)-(3,7,11b-triaza-benzo[c]fluoren-6-yl)-methanone-   (4-Methyl-piperazin-1-yl)-(3,7,11b-triaza-benzo[c]fluoren-6-yl)-methanone-   (3,5-Dimethyl-piperazin-1-yl)-(3,7,11b-triaza-benzo[c]fluoren-6-yl)-methanone-   6-(5-Methyl-[1,3,4]oxadiazol-2-yl)-3,7,11b-triaza-benzo[c]fluorine-   6-(4H-[1,2,4]Triazol-3-yl)-3,7,11b-triaza-benzo[c]fluorine-   2-Methyl-3,7,11b-triaza-benzo[c]fluorene-6-carboxylic acid    (2-pyrrolidin-1-yl-ethyl)-amide-   2,4-Dimethyl-3,7,11b-triaza-benzo[c]fluorene-6-carboxylic acid    (2-pyrrolidin-1-yl-ethyl)-amide-   2-Methyl-3,7,11b-triaza-benzo[c]fluorene-6-carboxylic acid    (2-morpholin-4-yl-ethyl)-amide-   (4-Methyl-[1,4]diazepan-1-yl)-(2-methyl-3,7,11b-triaza-benzo[c]fluoren-6-yl)-methanone

A method for preparing compounds of formula II(A), III(A), VII(A),VII(B) VIII(A), VIII(B) and VIII(C) are illustrated in Exemplary Scheme17.

Compound 51 is prepared according to U.S. Pat. No. 7,816,524. Thechlorination of compound 51 using chlorinating agent such as POCl₃ leadsto compound 52. Compound 52 undergoes nucleophilic substitution withsubstituted amines to yield 53. Compound 54 is obtained by treatingcompound 53 by substituted amine or by forming first carboxylic acid viahydrolysis of the ester group then treating with substituted amines toperform the amide coupling reaction.

Compounds described herein are also prepared according to ExemplaryScheme 15.

Non-limiting specific examples that can be prepared by the method setforth in Exemplary Scheme 17 include:

Certain Terminology

Unless otherwise stated, the following terms used in this applicationhave the definitions given below. The use of the term “including” aswell as other forms, such as “include”, “includes,” and “included,” isnot limiting. The section headings used herein are for organizationalpurposes only and are not to be construed as limiting the subject matterdescribed.

As used herein, C₁-C_(x) includes C₁-C₂, C₁-C₃ . . . C₁-C_(x). By way ofexample only, a group designated as “C₁-C₄” indicates that there are oneto four carbon atoms in the moiety, i.e. groups containing 1 carbonatom, 2 carbon atoms, 3 carbon atoms or 4 carbon atoms. Thus, by way ofexample only, “C₁-C₄ alkyl” indicates that there are one to four carbonatoms in the alkyl group, i.e., the alkyl group is selected from amongmethyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, andt-butyl.

An “alkyl” group refers to an aliphatic hydrocarbon group. The alkylgroup is branched or straight chain. In some embodiments, the “alkyl”group has 1 to 10 carbon atoms, i.e. a C₁-C₁₀alkyl. Whenever it appearsherein, a numerical range such as “1 to 10” refers to each integer inthe given range; e.g., “1 to 10 carbon atoms” means that the alkyl groupconsist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up toand including 10 carbon atoms, although the present definition alsocovers the occurrence of the term “alkyl” where no numerical range isdesignated. In some embodiments, an alkyl is a C₁-C₆alkyl. In one aspectthe alkyl is methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl,sec-butyl, or t-butyl. Typical alkyl groups include, but are in no waylimited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl,sec-butyl, tertiary butyl, pentyl, neopentyl, or hexyl.

An “alkylene” group refers to a divalent alkyl radical. Any of the abovementioned monovalent alkyl groups may be an alkylene by abstraction of asecond hydrogen atom from the alkyl. In some embodiments, an alkelene isa C₁-C₆alkylene. In other embodiments, an alkylene is a C₁-C₄alkylene.Typical alkylene groups include, but are not limited to, —CH₂—,—CH(CH₃)—, —C(CH₃)₂—, —CH₂CH₂—, —CH₂CH(CH₃)—, —CH₂C(CH₃)₂—, —CH₂CH₂CH₂—,—CH₂CH₂CH₂CH₂—, and the like.

“Azacyclic” or “azacyclic ring” refers to a saturated, partiallyunsaturated, or aromatic 3-7 membered monocyclic ring or an 8-12membered fused bicyclic ring system containing at least one nitrogenatom. Such azacyclic rings may optionally contain from 1-2 additionalheteroatoms selected from N, O, and S as ring members, and mayoptionally be substituted to the extent such substitutions make chemicalsense.

“Deuteroalkyl” refers to an alkyl group where 1 or more hydrogen atomsof an alkyl are replaced with deuterium.

The term “alkenyl” refers to a type of alkyl group in which at least onecarbon-carbon double bond is present. In one embodiment, an alkenylgroup has the formula —C(R)═CR₂, wherein R refers to the remainingportions of the alkenyl group, which may be the same or different. Insome embodiments, R is H or an alkyl. Non-limiting examples of analkenyl group include —CH═CH₂, —C(CH₃)═CH₂, —CH═CHCH₃, —C(CH₃)═CHCH₃,and —CH₂CH═CH₂.

The term “alkynyl” refers to a type of alkyl group in which at least onecarbon-carbon triple bond is present. In one embodiment, an alkenylgroup has the formula —C≡C—R, wherein R refers to the remaining portionsof the alkynyl group. In some embodiments, R is H or an alkyl.Non-limiting examples of an alkynyl group include —C≡CH,—C≡CCH₃—C≡CCH₂CH₃, —CH₂C≡CH.

An “alkoxy” group refers to a (alkyl)O— group, where alkyl is as definedherein.

The term “alkylamine” refers to the —N(alkyl)_(x)H_(y) group, where x is0 and y is 2, or where x is 1 and y is 1, or where x is 2 and y is 0.

The term “aromatic” refers to a planar ring having a delocalizedπ-electron system containing 4n+2π electrons, where n is an integer. Theterm “aromatic” includes both carbocyclic aryl (“aryl”, e.g., phenyl)and heterocyclic aryl (or “heteroaryl” or “heteroaromatic”) groups(e.g., pyridine). The term includes monocyclic or fused-ring polycyclic(i.e., rings which share adjacent pairs of carbon atoms) groups.

The term “carbocyclic” or “carbocycle” refers to a ring or ring systemwhere the atoms forming the backbone of the ring are all carbon atoms.The term thus distinguishes carbocyclic from “heterocyclic” rings or“heterocycles” in which the ring backbone contains at least one atomwhich is different from carbon. In some embodiments, at least one of thetwo rings of a bicyclic carbocycle is aromatic. In some embodiments,both rings of a bicyclic carbocycle are aromatic.

As used herein, the term “aryl” refers to an aromatic ring wherein eachof the atoms forming the ring is a carbon atom. In one aspect, aryl isphenyl or a naphthyl. In some embodiments, an aryl is a phenyl. In someembodiments, an aryl is a C₆-C₁₀aryl. Depending on the structure, anaryl group is a monoradical or a diradical (i.e., an arylene group).

The term “cycloalkyl” refers to a monocyclic or polycyclic aliphatic,non-aromatic radical, wherein each of the atoms forming the ring (i.e.skeletal atoms) is a carbon atom. In some embodiments, cycloalkyls arespirocyclic or bridged compounds. In some embodiments, cycloalkyls areoptionally fused with an aromatic ring, and the point of attachment isat a carbon that is not an aromatic ring carbon atom. Cycloalkyl groupsinclude groups having from 3 to 10 ring atoms. In some embodiments,cycloalkyl groups are selected from among cyclopropyl, cyclobutyl,cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl,cyclooctyl, spiro[2.2]pentyl, norbornyl and bicycle[1.1.1]pentyl. Insome embodiments, a cycloalkyl is a C₃-C₆cycloalkyl.

The term “halo” or, alternatively, “halogen” or “halide” means fluoro,chloro, bromo or iodo. In some embodiments, halo is fluoro, chloro, orbromo.

The term “fluoroalkyl” refers to an alkyl in which one or more hydrogenatoms are replaced by a fluorine atom. In one aspect, a fluoroalkyl is aC₁-C₆ fluoroalkyl.

The term “heteroalkyl” refers to an alkyl group in which one or moreskeletal atoms of the alkyl are selected from an atom other than carbon,e.g., oxygen, nitrogen (e.g. —NH—, —N(alkyl)-, sulfur, or combinationsthereof. A heteroalkyl is attached to the rest of the molecule at acarbon atom of the heteroalkyl. In one aspect, a heteroalkyl is aC₁-C₆heteroalkyl.

The term “heterocycle” or “heterocyclic” refers to heteroaromatic rings(also known as heteroaryls) and heterocycloalkyl rings (also known asheteroalicyclic groups) containing one to four heteroatoms in thering(s), where each heteroatom in the ring(s) is selected from O, S andN, wherein each heterocyclic group has from 3 to 10 atoms in its ringsystem, and with the proviso that any ring does not contain two adjacentO or S atoms. Non-aromatic heterocyclic groups (also known asheterocycloalkyls) include rings having 3 to 10 atoms in its ring systemand aromatic heterocyclic groups include rings having 5 to 10 atoms inits ring system. The heterocyclic groups include benzo-fused ringsystems. Examples of non-aromatic heterocyclic groups are pyrrolidinyl,tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, oxazolidinonyl,tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidinyl,morpholinyl, thiomorpholinyl, thioxanyl, piperazinyl, aziridinyl,azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl,oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl,pyrrolin-2-yl, pyrrolin-3-yl, indolinyl, 2H-pyranyl, 4H-pyranyl,dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl,dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl,imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl,3-azabicyclo[4.1.0]heptanyl, 3H-indolyl, indolin-2-onyl,isoindolin-1-onyl, isoindoline-1,3-dionyl,3,4-dihydroisoquinolin-1(2H)-onyl, 3,4-dihydroquinolin-2(1H)-onyl,isoindoline-1,3-dithionyl, benzo[d]oxazol-2(3H)-onyl,1H-benzo[d]imidazol-2(3H)-onyl, benzo[d]thiazol-2(3H)-onyl, andquinolizinyl. Examples of aromatic heterocyclic groups are pyridinyl,imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl,furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl,quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl,cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl,triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl,furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl,benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, andfuropyridinyl. The foregoing groups are either C-attached (or C-linked)or N-attached where such is possible. For instance, a group derived frompyrrole includes both pyrrol-1-yl (N-attached) or pyrrol-3-yl(C-attached). Further, a group derived from imidazole includesimidazol-1-yl or imidazol-3-yl (both N-attached) or imidazol-2-yl,imidazol-4-yl or imidazol-5-yl (all C-attached). The heterocyclic groupsinclude benzo-fused ring systems. Non-aromatic heterocycles areoptionally substituted with one or two oxo (═O) moieties, such aspyrrolidin-2-one. In some embodiments, at least one of the two rings ofa bicyclic heterocycle is aromatic. In some embodiments, both rings of abicyclic heterocycle are aromatic.

The terms “heteroaryl” or, alternatively, “heteroaromatic” refers to anaryl group that includes one or more ring heteroatoms selected fromnitrogen, oxygen and sulfur. Illustrative examples of heteroaryl groupsinclude monocyclic heteroaryls and bicyclic heteroaryls. Monocyclicheteroaryls include pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl,triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl,oxazolyl, isothiazolyl, pyrrolyl, pyridazinyl, triazinyl, oxadiazolyl,thiadiazolyl, and furazanyl. Bicyclic heteroaryls include indolizine,indole, benzofuran, benzothiophene, indazole, benzimidazole, purine,quinolizine, quinoline, isoquinoline, cinnoline, phthalazine,quinazoline, quinoxaline, 1,8-naphthyridine, and pteridine. In someembodiments, a heteroaryl contains 0-4 N atoms in the ring. In someembodiments, a heteroaryl contains 1-4 N atoms in the ring. In someembodiments, a heteroaryl contains 0-4 N atoms, 0-1 O atoms, and 0-1 Satoms in the ring. In some embodiments, a heteroaryl contains 1-4 Natoms, 0-1 O atoms, and 0-1 S atoms in the ring. In some embodiments,heteroaryl is a C₁-C₉heteroaryl. In some embodiments, monocyclicheteroaryl is a C₁-C₅heteroaryl. In some embodiments, monocyclicheteroaryl is a 5-membered or 6-membered heteroaryl. In someembodiments, bicyclic heteroaryl is a C₆-C₉heteroaryl.

A “heterocycloalkyl” or “heteroalicyclic” group refers to a cycloalkylgroup that includes at least one heteroatom selected from nitrogen,oxygen and sulfur. In some embodiments, a heterocycloalkyl is fused withan aryl or heteroaryl. In some embodiments, the heterocycloalkyl isoxazolidinonyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl,tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl,thiomorpholinyl, piperazinyl, piperidin-2-onyl,pyrrolidine-2,5-dithionyl, pyrrolidine-2,5-dionyl, pyrrolidinonyl,imidazolidinyl, imidazolidin-2-onyl, or thiazolidin-2-onyl. The termheteroalicyclic also includes all ring forms of the carbohydrates,including but not limited to the monosaccharides, the disaccharides andthe oligosaccharides. In one aspect, a heterocycloalkyl is aC₂-C₁₀heterocycloalkyl. In another aspect, a heterocycloalkyl is aC₄-C₁₀heterocycloalkyl. In some embodiments, a heterocycloalkyl contains0-2 N atoms in the ring. In some embodiments, a heterocycloalkylcontains 0-2 N atoms, 0-2 O atoms and 0-1 S atoms in the ring.

The term “bond” or “single bond” refers to a chemical bond between twoatoms, or two moieties when the atoms joined by the bond are consideredto be part of larger substructure. In one aspect, when a group describedherein is a bond, the referenced group is absent thereby allowing a bondto be formed between the remaining identified groups.

The term “moiety” refers to a specific segment or functional group of amolecule. Chemical moieties are often recognized chemical entitiesembedded in or appended to a molecule.

The term “optionally substituted” or “substituted” means that thereferenced group is optionally substituted with one or more additionalgroup(s) individually and independently selected from halogen, —CN,—NH₂, —NH(alkyl), —N(alkyl)₂, —OH, —CO₂H, —CO₂alkyl, —C(═O)NH₂,—C(═O)NH(alkyl), —C(═O)N(alkyl)₂, —S(═O)₂NH₂, —S(═O)₂NH(alkyl),—S(═O)₂N(alkyl)₂, alkyl, cycloalkyl, fluoroalkyl, heteroalkyl, alkoxy,fluoroalkoxy, heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio,arylthio, alkylsulfoxide, arylsulfoxide, alkylsulfone, and arylsulfone.In some other embodiments, optional substituents are independentlyselected from halogen, —CN, —NH₂, —NH(CH₃), —N(CH₃)₂, —OH, —CO₂H,—CO₂(C₁-C₄alkyl), —C(═O)NH₂, —C(═O)NH(C₁-C₄alkyl), —C(═O)N(C₁-C₄alkyl)₂,—S(═O)₂NH₂, —S(═O)₂NH(C₁-C₄alkyl), —S(═O)₂N(C₁-C₄alkyl)₂, C₁-C₄alkyl,C₃-C₆cycloalkyl, C₁-C₄fluoroalkyl, C₁-C₄heteroalkyl, C₁-C₄alkoxy,C₁-C₄fluoroalkoxy, —SC₁-C₄alkyl, —S(═O)C₁-C₄alkyl, and—S(═O)₂C₁-C₄alkyl. In some embodiments, optional substituents areindependently selected from halogen, —CN, —NH₂, —OH, —NH(CH₃), —N(CH₃)₂,—CH₃, —CH₂CH₃, —CF₃, —OCH₃, and —OCF₃. In some embodiments, substitutedgroups are substituted with one or two of the preceding groups. In someembodiments, an optional substituent on an aliphatic carbon atom(acyclic or cyclic) includes oxo (═O).

“Carboxylate bioisostere” or “carboxy bioisostere” as used herein refersto a moiety that is expected to be negatively charged to a substantialdegree at physiological pH. In certain embodiments, the carboxylatebioisostere is a moiety selected from the group consisting of:

and salts of the forgoing, wherein each R″ is independently H or anoptionally substituted member selected from the group consisting ofC₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ heteroalkyl, C₃₋₈ carbocyclic ring; orR″ is a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀ heteroalkyl substitutedwith an optionally substituted C₃₋₈ carbocyclic ring or C₃₋₈heterocyclic ring.

Amide bioisostere and ester bioisostere as used herein refer to amoieties represented by the following examples:

wherein each R″ is independently H or an optionally substituted memberselected from the group consisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀heteroalkyl, C₃₋₈ carbocyclic ring; or R″ is a C₁₋₁₀ alkyl, C₂₋₁₀alkenyl, or C₂₋₁₀ heteroalkyl substituted with an optionally substitutedC₃₋₈ carbocyclic ring or C₃₋₈ heterocyclic ring.

The compounds presented herein may possess one or more stereocenters andeach center may exist in the R or S configuration. The compoundspresented herein include all diastereomeric, enantiomeric, and epimericforms as well as the appropriate mixtures thereof. Stereoisomers may beobtained, if desired, by methods known in the art such as, for example,the separation of individual stereoisomers by chiral chromatographiccolumns or by stereoselective synthesis.

A “quaternary amine” is a positively charged polyatomic ion of thestructure NR₄ ⁺, where R is an alkyl or aryl group. The four (4) Rgroups that make up the quaternary amine may be the same or differentand may be connected to one another. Quaternary amines can be preparedby the alkylation of tertiary amines, in a process calledquaternization, as well as by other methods known in the art.

The term “acceptable” with respect to a formulation, composition oringredient, as used herein, means having no persistent detrimentaleffect on the general health of the subject being treated.

The term “modulate” as used herein, means to interact with a targeteither directly or indirectly so as to alter the activity of the target,including, by way of example only, to enhance the activity of thetarget, to inhibit the activity of the target, to limit the activity ofthe target, or to extend the activity of the target.

The term “modulator” as used herein, refers to a molecule that interactswith a target either directly or indirectly. The interactions include,but are not limited to, the interactions of an agonist, partial agonist,an inverse agonist, antagonist, degrader, or combinations thereof. Insome embodiments, a modulator is an antagonist. In some embodiments, amodulator is a degrader.

The terms “administer,” “administering”, “administration,” and the like,as used herein, refer to the methods that may be used to enable deliveryof compounds or compositions to the desired site of biological action.These methods include, but are not limited to oral routes, intraduodenalroutes, parenteral injection (including intravenous, subcutaneous,intraperitoneal, intramuscular, intravascular or infusion), topical andrectal administration. Those of skill in the art are familiar withadministration techniques that can be employed with the compounds andmethods described herein. In some embodiments, the compounds andcompositions described herein are administered orally.

The terms “co-administration” or the like, as used herein, are meant toencompass administration of the selected therapeutic agents to a singlepatient, and are intended to include treatment regimens in which theagents are administered by the same or different route of administrationor at the same or different time.

The terms “effective amount” or “therapeutically effective amount,” asused herein, refer to a sufficient amount of an agent or a compoundbeing administered, which will relieve to some extent one or more of thesymptoms of the disease or condition being treated. The result includesreduction and/or alleviation of the signs, symptoms, or causes of adisease, or any other desired alteration of a biological system. Forexample, an “effective amount” for therapeutic uses is the amount of thecomposition comprising a compound as disclosed herein required toprovide a clinically significant decrease in disease symptoms. Anappropriate “effective” amount in any individual case is optionallydetermined using techniques, such as a dose escalation study.

The terms “enhance” or “enhancing,” as used herein, means to increase orprolong either in potency or duration a desired effect. Thus, in regardto enhancing the effect of therapeutic agents, the term “enhancing”refers to the ability to increase or prolong, either in potency orduration, the effect of other therapeutic agents on a system. An“enhancing-effective amount,” as used herein, refers to an amountadequate to enhance the effect of another therapeutic agent in a desiredsystem.

The term “pharmaceutical combination” as used herein, means a productthat results from the mixing or combining of more than one activeingredient and includes both fixed and non-fixed combinations of theactive ingredients. The term “fixed combination” means that the activeingredients, e.g. a compound described herein, or a pharmaceuticallyacceptable salt thereof, and a co-agent, are both administered to apatient simultaneously in the form of a single entity or dosage. Theterm “non-fixed combination” means that the active ingredients, e.g. acompound described herein, or a pharmaceutically acceptable saltthereof, and a co-agent, are administered to a patient as separateentities either simultaneously, concurrently or sequentially with nospecific intervening time limits, wherein such administration provideseffective levels of the two compounds in the body of the patient. Thelatter also applies to cocktail therapy, e.g. the administration ofthree or more active ingredients.

The terms “kit” and “article of manufacture” are used as synonyms.

The term “subject” or “patient” encompasses mammals. Examples of mammalsinclude, but are not limited to, any member of the Mammalian class:humans, non-human primates such as chimpanzees, and other apes andmonkey species; farm animals such as cattle, horses, sheep, goats,swine; domestic animals such as rabbits, dogs, and cats; laboratoryanimals including rodents, such as rats, mice and guinea pigs, and thelike. In one aspect, the mammal is a human.

The terms “treat,” “treating” or “treatment,” as used herein, includealleviating, abating or ameliorating at least one symptom of a diseaseor condition, preventing additional symptoms, inhibiting the disease orcondition, e.g., arresting the development of the disease or condition,relieving the disease or condition, causing regression of the disease orcondition, relieving a condition caused by the disease or condition, orstopping the symptoms of the disease or condition eitherprophylactically and/or therapeutically.

Pharmaceutical Compositions

In some embodiments, the compounds described herein are formulated intopharmaceutical compositions. Pharmaceutical compositions are formulatedin a conventional manner using one or more pharmaceutically acceptableinactive ingredients that facilitate processing of the active compoundsinto preparations that are used pharmaceutically. It is understood inthe art that proper formulation is dependent upon the route ofadministration chosen. A summary of pharmaceutical compositionsdescribed herein is found, for example, in Remington: The Science andPractice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack PublishingCompany, 1995); Hoover, John E., Remington's Pharmaceutical Sciences,Mack Publishing Co., Easton, Pa. 1975; Liberman, H. A. and Lachman, L.,Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980;and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed.(Lippincott Williams & Wilkins 1999), herein incorporated by referencefor such disclosure.

In some embodiments, the compounds described herein are administeredeither alone or in combination with pharmaceutically acceptablecarriers, excipients or diluents, in a pharmaceutical composition.Administration of the compounds and compositions described herein can beeffected by any method that enables delivery of the compounds to thesite of action. These methods include, though are not limited todelivery via enteral routes (including oral, gastric or duodenal feedingtube, rectal suppository and rectal enema), parenteral routes (injectionor infusion, including intraarterial, intracardiac, intradermal,intraduodenal, intramedullary, intramuscular, intraosseous,intraperitoneal, intrathecal, intravascular, intravenous, intravitreal,epidural and subcutaneous), inhalational, transdermal, transmucosal,sublingual, buccal and topical (including epicutaneous, dermal, enema,eye drops, ear drops, intranasal, vaginal) administration, although themost suitable route may depend upon for example the condition anddisorder of the recipient. By way of example only, compounds describedherein can be administered locally to the area in need of treatment, byfor example, local infusion during surgery, topical application such ascreams or ointments, injection, catheter, or implant. The administrationcan also be by direct injection at the site of a diseased tissue ororgan.

In some embodiments, pharmaceutical compositions suitable for oraladministration are presented as discrete units such as capsules, cachetsor tablets each containing a predetermined amount of the activeingredient; as a powder or granules; as a solution or a suspension in anaqueous liquid or a non-aqueous liquid; or as an oil-in-water liquidemulsion or a water-in-oil liquid emulsion. In some embodiments, theactive ingredient is presented as a bolus, electuary or paste.

Pharmaceutical compositions which can be used orally include tablets,push-fit capsules made of gelatin, as well as soft, sealed capsules madeof gelatin and a plasticizer, such as glycerol or sorbitol. Tablets maybe made by compression or molding, optionally with one or more accessoryingredients. Compressed tablets may be prepared by compressing in asuitable machine the active ingredient in a free-flowing form such as apowder or granules, optionally mixed with binders, inert diluents, orlubricating, surface active or dispersing agents. Molded tablets may bemade by molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent. In some embodiments, the tabletsare coated or scored and are formulated so as to provide slow orcontrolled release of the active ingredient therein. All formulationsfor oral administration should be in dosages suitable for suchadministration. The push-fit capsules can contain the active ingredientsin admixture with filler such as lactose, binders such as starches,and/or lubricants such as talc or magnesium stearate and, optionally,stabilizers. In soft capsules, the active compounds may be dissolved orsuspended in suitable liquids, such as fatty oils, liquid paraffin, orliquid polyethylene glycols. In some embodiments, stabilizers are added.Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or Dragee coatings for identification or to characterizedifferent combinations of active compound doses.

In some embodiments, pharmaceutical compositions are formulated forparenteral administration by injection, e.g., by bolus injection orcontinuous infusion. Formulations for injection may be presented in unitdosage form, e.g., in ampoules or in multi-dose containers, with anadded preservative. The compositions may take such forms as suspensions,solutions or emulsions in oily or aqueous vehicles, and may containformulatory agents such as suspending, stabilizing and/or dispersingagents. The compositions may be presented in unit-dose or multi-dosecontainers, for example sealed ampoules and vials, and may be stored inpowder form or in a freeze-dried (lyophilized) condition requiring onlythe addition of the sterile liquid carrier, for example, saline orsterile pyrogen-free water, immediately prior to use. Extemporaneousinjection solutions and suspensions may be prepared from sterilepowders, granules and tablets of the kind previously described.

Pharmaceutical compositions for parenteral administration includeaqueous and non-aqueous (oily) sterile injection solutions of the activecompounds which may contain antioxidants, buffers, bacteriostats andsolutes which render the formulation isotonic with the blood of theintended recipient; and aqueous and non-aqueous sterile suspensionswhich may include suspending agents and thickening agents. Suitablelipophilic solvents or vehicles include fatty oils such as sesame oil,or synthetic fatty acid esters, such as ethyl oleate or triglycerides,or liposomes. Aqueous injection suspensions may contain substances whichincrease the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may alsocontain suitable stabilizers or agents which increase the solubility ofthe compounds to allow for the preparation of highly concentratedsolutions.

Pharmaceutical compositions may also be formulated as a depotpreparation. Such long acting formulations may be administered byimplantation (for example subcutaneously or intramuscularly) or byintramuscular injection. Thus, for example, the compounds may beformulated with suitable polymeric or hydrophobic materials (forexample, as an emulsion in an acceptable oil) or ion exchange resins, oras sparingly soluble derivatives, for example, as a sparingly solublesalt.

For buccal or sublingual administration, the compositions may take theform of tablets, lozenges, pastilles, or gels formulated in conventionalmanner. Such compositions may comprise the active ingredient in aflavored basis such as sucrose and acacia or tragacanth.

Pharmaceutical compositions may also be formulated in rectalcompositions such as suppositories or retention enemas, e.g., containingconventional suppository bases such as cocoa butter, polyethyleneglycol, or other glycerides.

Pharmaceutical compositions may be administered topically, that is bynon-systemic administration. This includes the application of a compoundof the present invention externally to the epidermis or the buccalcavity and the instillation of such a compound into the ear, eye andnose, such that the compound does not significantly enter the bloodstream. In contrast, systemic administration refers to oral,intravenous, intraperitoneal and intramuscular administration.

Pharmaceutical compositions suitable for topical administration includeliquid or semi-liquid preparations suitable for penetration through theskin to the site of inflammation such as gels, liniments, lotions,creams, ointments or pastes, and drops suitable for administration tothe eye, ear or nose. The active ingredient may comprise, for topicaladministration, from 0.001% to 10% w/w, for instance from 1% to 2% byweight of the formulation.

Pharmaceutical compositions for administration by inhalation areconveniently delivered from an insufflator, nebulizer pressurized packsor other convenient means of delivering an aerosol spray. Pressurizedpacks may comprise a suitable propellant such asdichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In thecase of a pressurized aerosol, the dosage unit may be determined byproviding a valve to deliver a metered amount. Alternatively, foradministration by inhalation or insufflation, pharmaceuticalpreparations may take the form of a dry powder composition, for examplea powder mix of the compound and a suitable powder base such as lactoseor starch. The powder composition may be presented in unit dosage form,in for example, capsules, cartridges, gelatin or blister packs fromwhich the powder may be administered with the aid of an inhalator orinsufflator.

It should be understood that in addition to the ingredients particularlymentioned above, the compounds and compositions described herein mayinclude other agents conventional in the art having regard to the typeof formulation in question, for example those suitable for oraladministration may include flavoring agents.

Methods of Dosing and Treatment Regimens

In one embodiment, the compounds described herein, or a pharmaceuticallyacceptable salt thereof, are used in the preparation of medicaments forthe treatment of diseases or conditions in a mammal that would benefitfrom inhibiting POL1 transcription. Methods for treating any of thediseases or conditions described herein in a mammal in need of suchtreatment, involves administration of pharmaceutical compositions thatinclude at least one compound described herein or a pharmaceuticallyacceptable salt, active metabolite, prodrug, or pharmaceuticallyacceptable solvate thereof, in therapeutically effective amounts to saidmammal.

In some embodiments, the invention provides methods of treatingconditions associated with polymerase I transcription, comprising:administering to a patient in need thereof an effective amount of acompound of the invention. In another embodiment, the invention providesa method of inhibiting polymerase I transcription: comprising,contacting the enzyme with a compound of the invention. In a furtherembodiment, the invention provides a method of inhibiting polymerase Itranscription: comprising, administering a first compound to a subjectthat is converted in vivo to a compound of the invention.

“Conditions associated with polymerase I transcription” includedisorders and diseases in which the inhibition of polymerase Itranscription provides a therapeutic benefit, such as cancer,allergy/asthma, diseases and conditions of the immune system,inflammation, disease and conditions of the central nervous system(CNS), cardiovascular disease, viral infections, dermatological disease,and diseases and conditions related to uncontrolled angiogenesis, andthe like. Where general terms are used herein to describe conditionsassociated with polymerase I transcription it is understood that themore specifically described conditions mentioned in the variousdiagnostic manuals and other materials are included within the scope ofthis invention.

The term “cancer,” as used herein refers to an abnormal growth of cells,which tend to proliferate in an uncontrolled way and, in some cases, tometastasize (spread). The types of cancer include, but is not limitedto, solid tumors (such as those of the bladder, bowel, brain, breast,endometrium, heart, kidney, lung, lymphatic tissue (lymphoma), ovary,pancreas or other endocrine organ (thyroid), prostate, skin (melanoma)or hematological tumors (such as the leukemias). See, Ding X Z et al.,Anticancer Drugs. 2005 June; 16(5):467-73. Review; Chen X et al., ClinCancer Res. 2004 Oct. 1; 10(19):6703-9, each of which are incorporatedby reference herein in their entirety.

For example, it is understood that the treatment of cancer includestreatment of all neoplasia, regardless of their histopathologicalappearance. Particularly, the cancers that can be treated include, butare not limited to, cancer of blood, including myelofibrosis, leukemia(including acute myelogenous leukemia, chronic myelogenous leukemia,acute lymphocytic leukemia, chronic lymphocytic leukemia), cancer of theskin, including melanoma, basal cell carcinoma, and squamous cellcarcinoma, bone, liver, lung (including small-cell lung tumor, nonsmall-cell lung cancer and bronchioalveolar cancer), brain, breast,prostate, larynx, gall bladder, pancreas, rectum, bile duct,parathyroid, thyroid, adrenal, neural tissue, bladder, spleen, head andneck, included the jaw, mouth, and nose, colon, stomach, testes,esophagus, uterus, cervix and vulva, colorectal, bronchi, bile duct,bladder, kidney, ovary, pancreas, multiple myeloma, lymphomas, basalcell carcinoma, squamous cell carcinoma of both ulcerating and papillarytype, osteo sarcoma, Ewing's sarcoma, veticulum cell sarcoma, myeloma,giant cell tumor, islet cell tumor, acute and chronic lymphocytic andgranulocytic tumors, hairy-cell tumor, adenoma, hyperplasia, medullarycarcinoma, pheochromocytoma, mucosal neuronms, intestinalganglioneuromas, hyperplastic corneal nerve tumor, marfanoid habitustumor, Wilm's tumor, seminoma, ovarian tumor, leiomyomater tumor,cervical dysplasia and in situ carcinoma, neuroblastoma, retinoblastoma,myelodysplastic syndrome, mycosis fungicide, rhabdomyosarcoma,astrocytoma, non-Hodgkin's lymphoma, Kaposi's sarcoma, osteogenic andother sarcoma, malignant hypercalcemia, polycythermia vera,adenocarcinoma, glioblastoma multiforma, glioma, lymphomas, epidermoidcarcinomas, and other carcinomas and sarcomas.

Benign tumors may also be treated by the compounds of the presentinvention and include, but are not limited to, hemangiomas,hepatocellular adenoma, cavernous haemangioma, focal nodularhyperplasia, acoustic neuromas, neurofibroma, bile duct adenoma, bileduct cystanoma, fibroma, lipomas, leiomyomas, mesotheliomas, teratomas,myxomas, nodular regenerative hyperplasia, trachomas, pyogenicgranulomas, and the like, and hamartoma conditions such as Peutz-JeghersSyndrome (PJS), Cowden disease, Bannayan-Riley-Ruvalcaba Syndrome(BRRS), Proteus syndrome, Lhermitte-Duclos disease and TuberousSclerosis (TSC).

The compounds of the present invention may also be used to treatabnormal cell proliferation due to insults to body tissue duringsurgery. These insults may arise as a result of a variety of surgicalprocedures such as joint surgery, bowel surgery, and cheloid scarring.Diseases that produce fibrotic tissue include emphysema. Repetitivemotion disorders that may be treated using the present invention includecarpal tunnel syndrome.

The compounds of the invention may also be useful in the prevention ofrestenosis that is the control of undesired proliferation of normalcells in the vasculature in response to the introduction of stents inthe treatment of vasculature disease.

Proliferative responses associated with organ transplantation that maybe treated using Pol I transcription inhibitors of the invention includeproliferative responses contributing to potential organ rejections orassociated complications. Specifically, these proliferative responsesmay occur during transplantation of the heart, lung, liver, kidney, andother body organs or organ systems.

The compounds of the invention may also be useful the treatment ofabnormal angiogenesis including the abnormal angiogenesis accompanyingrheumatoid arthritis, ischemic-reperfusion related brain edema andinjury, cortical ischemia, ovarian hyperplasia and hypervascularity,(polycystic ovary syndrome), endometriosis, psoriasis, diabeticretinopaphy, and other ocular angiogenic diseases such as retinopathy ofprematurity (retrolental fibroplastic), macular degeneration, cornealgraft rejection, neuroscular glaucoma, Oster Webber syndrome,retinal/choroidal neuvascularization and corneal neovascularization,Best's disease, myopia, optic pits, Stargart's diseases, Pagets disease,vein occlusion, artery occlusion, sickle cell anemia, sarcoid, syphilis,pseudoxanthoma elasticum carotid abstructive diseases, chronicuveitis/vitritis, mycobacterial infections, Lyme's disease, systemiclupus erythematosis, retinopathy of prematurity, Eales disease, diabeticretinopathy, macular degeneration, Bechets diseases, infections causinga retinitis or chroiditis, presumed ocular histoplasmosis, parsplanitis, chronic retinal detachment, hyperviscosity syndromes,toxoplasmosis, trauma and post-laser complications, diseases associatedwith rubesis (neovascularization of the angle), diseases caused by theabnormal proliferation of fibrovascular or fibrous tissue including allforms of proliferative vitreoretinopathy, atopic keratitis, superiorlimbic keratitis, pterygium keratitis sicca, sjogrens, acne rosacea,phylectenulosis, diabetic retinopathy, retinopathy of prematurity,corneal graft rejection, Mooren's ulcer, Terrien's marginaldegeneration, marginal keratolysis, polyarteritis, Wegener sarcoidosis,scleritis, periphigoid radial keratotomy, neovascular glaucoma andretrolental fibroplasia, syphilis, Mycobacteria infections, lipiddegeneration, chemical burns, bacterial ulcers, fungal ulcers, Herpessimplex infections, Herpes zoster infections, protozoan infections, andKaposi sarcoma, Alzheimer's disease, Parkinson's disease amyotrophiclateral sclerosis (ALS), epilepsy, seizures, Huntington's disease,polyglutamine diseases, traumatic brain injury, ischemic andhemorrhaging stroke, cerebral ischemias or neurodegenerative disease,including apoptosis-driven neurodegenerative disease, caused bytraumatic injury, acute hypoxia, ischemia or glutamate neurotoxicity.

For example, it is understood that treatments of inflammation include,but are not limited to, acute pancreatitis, chronic pancreatitis,asthma, allergies, chronic obstructive pulmonary disease, adultrespiratory distress syndrome and chronic inflammatory diseasesassociated with uncontrolled angiogenesis, inflammatory bowel diseasessuch as Crohn's disease and ulcerative colitis, psoriasis, sarcoidois,and rheumatoid arthritis, sarcoidosis, and multisystem granulomatousdisorder.

For example, it is understood that treatment of autoimmune includes, butis not limited to, glomerulonephritis, rheumatoid arthritis, systemiclupus erythematosus, scleroderma, chronic thyroiditis, Graves' disease,autoimmune gastritis, diabetes, autoimmune hemolytic anemia, autoimmuneneutropenia, thrombocytopenia, atopic dermatitis, chronic activehepatitis, myasthenia gravis, multiple sclerosis, inflammatory boweldisease, ulcerative colitis, Crohn's disease, psoriasis, graft vs. hostdisease, multiple sclerosis, or Sjoegren's syndrome.

In certain embodiments, the compositions containing the compound(s)described herein are administered for prophylactic and/or therapeutictreatments. In certain therapeutic applications, the compositions areadministered to a patient already suffering from a disease or condition,in an amount sufficient to cure or at least partially arrest at leastone of the symptoms of the disease or condition. Amounts effective forthis use depend on the severity and course of the disease or condition,previous therapy, the patient's health status, weight, and response tothe drugs, and the judgment of the treating physician. Therapeuticallyeffective amounts are optionally determined by methods including, butnot limited to, a dose escalation and/or dose ranging clinical trial.

In prophylactic applications, compositions containing the compoundsdescribed herein are administered to a patient susceptible to orotherwise at risk of a particular disease, disorder or condition. Suchan amount is defined to be a “prophylactically effective amount ordose.” In this use, the precise amounts also depend on the patient'sstate of health, weight, and the like. When used in patients, effectiveamounts for this use will depend on the severity and course of thedisease, disorder or condition, previous therapy, the patient's healthstatus and response to the drugs, and the judgment of the treatingphysician. In one aspect, prophylactic treatments include administeringto a mammal, who previously experienced at least one symptom of thedisease being treated and is currently in remission, a pharmaceuticalcomposition comprising a compound described herein, or apharmaceutically acceptable salt thereof, in order to prevent a returnof the symptoms of the disease or condition.

In certain embodiments wherein the patient's condition does not improve,upon the doctor's discretion the administration of the compounds areadministered chronically, that is, for an extended period of time,including throughout the duration of the patient's life in order toameliorate or otherwise control or limit the symptoms of the patient'sdisease or condition.

In certain embodiments wherein a patient's status does improve, the doseof drug being administered is temporarily reduced or temporarilysuspended for a certain length of time (i.e., a “drug holiday”). Inspecific embodiments, the length of the drug holiday is between 2 daysand 1 year, including by way of example only, 2 days, 3 days, 4 days, 5days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, ormore than 28 days. The dose reduction during a drug holiday is, by wayof example only, by 10%-100%, including by way of example only 10%, 15%,20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95%, and 100%.

Once improvement of the patient's conditions has occurred, a maintenancedose is administered if necessary. Subsequently, in specificembodiments, the dosage or the frequency of administration, or both, isreduced, as a function of the symptoms, to a level at which the improveddisease, disorder or condition is retained. In certain embodiments,however, the patient requires intermittent treatment on a long-termbasis upon any recurrence of symptoms.

The amount of a given agent that corresponds to such an amount variesdepending upon factors such as the particular compound, diseasecondition and its severity, the identity (e.g., weight, sex) of thesubject or host in need of treatment, but nevertheless is determinedaccording to the particular circumstances surrounding the case,including, e.g., the specific agent being administered, the route ofadministration, the condition being treated, and the subject or hostbeing treated.

In general, however, doses employed for adult human treatment aretypically in the range of 0.01 mg-5000 mg per day. In one aspect, dosesemployed for adult human treatment are from about 1 mg to about 1000 mgper day. In one embodiment, the desired dose is conveniently presentedin a single dose or in divided doses administered simultaneously or atappropriate intervals, for example as two, three, four or more sub-dosesper day.

In one embodiment, the daily dosages appropriate for the compounddescribed herein, or a pharmaceutically acceptable salt thereof, arefrom about 0.01 to about 50 mg/kg per body weight. In some embodiments,the daily dosage or the amount of active in the dosage form are lower orhigher than the ranges indicated herein, based on a number of variablesin regard to an individual treatment regime. In various embodiments, thedaily and unit dosages are altered depending on a number of variablesincluding, but not limited to, the activity of the compound used, thedisease or condition to be treated, the mode of administration, therequirements of the individual subject, the severity of the disease orcondition being treated, and the judgment of the practitioner.

Toxicity and therapeutic efficacy of such therapeutic regimens aredetermined by standard pharmaceutical procedures in cell cultures orexperimental animals, including, but not limited to, the determinationof the LD₅₀ and the ED₅₀. The dose ratio between the toxic andtherapeutic effects is the therapeutic index and it is expressed as theratio between LD₅₀ and ED₅₀. In certain embodiments, the data obtainedfrom cell culture assays and animal studies are used in formulating thetherapeutically effective daily dosage range and/or the therapeuticallyeffective unit dosage amount for use in mammals, including humans. Insome embodiments, the daily dosage amount of the compounds describedherein lies within a range of circulating concentrations that includethe ED₅₀ with minimal toxicity. In certain embodiments, the daily dosagerange and/or the unit dosage amount varies within this range dependingupon the dosage form employed and the route of administration utilized.

In any of the aforementioned aspects are further embodiments in whichthe effective amount of the compound described herein, or apharmaceutically acceptable salt thereof, is: (a) systemicallyadministered to the mammal; and/or (b) administered orally to themammal; and/or (c) intravenously administered to the mammal; and/or (d)administered by injection to the mammal; and/or (e) administeredtopically to the mammal; and/or (f) administered non-systemically orlocally to the mammal.

In any of the aforementioned aspects are further embodiments comprisingsingle administrations of the effective amount of the compound,including further embodiments in which (i) the compound is administeredonce a day; or (ii) the compound is administered to the mammal multipletimes over the span of one day.

In any of the aforementioned aspects are further embodiments comprisingmultiple administrations of the effective amount of the compound,including further embodiments in which (i) the compound is administeredcontinuously or intermittently: as in a single dose; (ii) the timebetween multiple administrations is every 6 hours; (iii) the compound isadministered to the mammal every 8 hours; (iv) the compound isadministered to the mammal every 12 hours; (v) the compound isadministered to the mammal every 24 hours. In further or alternativeembodiments, the method comprises a drug holiday, wherein theadministration of the compound is temporarily suspended or the dose ofthe compound being administered is temporarily reduced; at the end ofthe drug holiday, dosing of the compound is resumed. In one embodiment,the length of the drug holiday varies from 2 days to 1 year.

In certain instances, it is appropriate to administer at least onecompound described herein, or a pharmaceutically acceptable saltthereof, in combination with one or more other therapeutic agents. Incertain embodiments, the pharmaceutical composition further comprisesone or more anti-cancer agents.

In one embodiment, the therapeutic effectiveness of one of the compoundsdescribed herein is enhanced by administration of an adjuvant (i.e., byitself the adjuvant has minimal therapeutic benefit, but in combinationwith another therapeutic agent, the overall therapeutic benefit to thepatient is enhanced). Or, in some embodiments, the benefit experiencedby a patient is increased by administering one of the compoundsdescribed herein with another agent (which also includes a therapeuticregimen) that also has therapeutic benefit.

A wide variety of therapeutic agents may have a therapeutic additive orsynergistic effect with the compounds according to the presentinvention. Combination therapies that comprise one or more compounds ofthe present invention with one or more other therapeutic agents can beused, for example, to: (1) enhance the therapeutic effect(s) of the oneor more compounds of the present invention and/or the one or more othertherapeutic agents; (2) reduce the side effects exhibited by the one ormore compounds of the present invention and/or the one or more othertherapeutic agents; and/or (3) reduce the effective dose of the one ormore compounds of the present invention and/or the one or more othertherapeutic agents. It is noted that combination therapy is intended tocover when agents are administered before or after each other(sequential therapy) as well as when the agents are administered at thesame time.

Examples of such therapeutic agents that may be used in combination withthe present compounds include, but are not limited to, anti-cellproliferation agents, anticancer agents, alkylating agents, antibioticagents, antimetabolic agents, hormonal agents, plant-derived agents, andbiologic agents.

Anti-cell proliferation agents useful in combination with the compoundsof the present invention include, but are not limited to, retinoid acidand derivatives thereof, 2-methoxyestradiol, ANGIOSTATIN™ protein,ENDOSTATIN™ protein, suramin, squalamine, tissue inhibitor ofmetalloproteinase-I, tissue inhibitor of metalloproteinase-2,plasminogen activator inhibitor-1, plasminogen activator inhibitor-2,cartilage-derived inhibitor, paclitaxel, platelet factor 4, protaminesulphate (clupeine), sulphated chitin derivatives (prepared from queencrab shells), sulphated polysaccharide peptidoglycan complex (sp-pg),staurosporine, modulators of matrix metabolism, including for example,proline analogs ((1-azetidine-2-carboxylic acid (LACA),cishydroxyproline, d,l-3,4-dehydroproline, thiaproline,beta-aminopropionitrile fumarate,4-propyl-5-(4-pyridinyl)-2(3H)-oxazolone, methotrexate, mitoxantrone,heparin, interferons, 2 macroglobulin-serum, chimp-3, chymostatin,beta.-cyclodextrin tetradecasulfate, eponemycin; fumagillin, gold sodiumthiomalate, d-penicillamine (CDPT), beta-1-anticollagenase-serum,alpha-2-antiplasmin, bisantrene, lobenzarit disodium,n-(2-carboxyphenyl-4-chloroanthronilic acid disodium or “CCA”,thalidomide, angostatic steroid, cargboxynaminolmidazole,metalloproteinase inhibitors such as BB94. Other anti-angiogenesisagents that may be used include antibodies, preferably monoclonalantibodies against these angiogenic growth factors: bFGF, aFGF, FGF-5,VEGF isoforms, VEGF-C, HGF/SF and Ang-1/Ang-2.

Inhibitors of mTOR, PI3K, MEK, MAPK, PIM or ERK kinases are useful incombination with the compounds of the present invention. Specifically,(R)-3-(2,3-Dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dioneuseful in combination with the compounds of the present invention.Inhibitors of Hedgehog kinase are useful in combination with thecompounds of the present invention. Proteasome inhibitors, in particularbortezomib is useful in combination with the compounds of the presentinvention.

NAE inhibitors, VPS34 inhibitors, Aurora kinase, including Aurora Ainhibitors, and EGFR inhibitors (both antibodies and kinase inhibitors)are useful in combination with the compounds of the present invention.

Alkylating agents useful in combination with the compounds disclosedherein include, but are not limited to, bischloroethylamines (nitrogenmustards, e.g. chlorambucil, cyclophosphamide, ifosfamide,mechlorethamine, melphalan, uracil mustard), aziridines (e.g. thiotepa),alkyl alkone sulfonates (e.g. busulfan), nitrosoureas (e.g. carmustine,lomustine, streptozocin), nonclassic alkylating agents (altretamine,dacarbazine, and procarbazine), platinum compounds (carboplastin andcisplatin). Combination therapy including a polymerase I inhibitor andan alkylating agent is expected to have therapeutic synergistic effectsin the treatment of cancer and reduce sides affects associated withthese chemotherapeutic agents.

Examples of antibiotic agents useful in combination with the compoundsdisclosed herein include, but are not limited to, anthracyclines (e.g.doxorubicin, daunorubicin, epirubicin, idarubicin and anthracenedione),mitomycin C, bleomycin, dactinomycin, plicatomycin. These antibioticagents interfere with cell growth by targeting different cellularcomponents.

Antimetabolic agents useful in combination with the compounds disclosedherein include, but are not limited to, fluorouracil (5-FU), floxuridine(5-FUdR), methotrexate, leucovorin, hydroxyurea, thioguanine (6-TG),mercaptopurine (6-MP), cytarabine, pentostatin, fludarabine phosphate,cladribine (2-CDA), asparaginase, and gemcitabine. Combination therapyincluding a compound disclosed herein and an antimetabolic agent isexpected to have therapeutic synergistic effects on cancer and reducesides affects associated with these chemotherapeutic agents.

Hormonal agents useful in combination with the compounds disclosedherein include synthetic estrogens (e.g. diethylstibestrol),antiestrogens (e.g. tamoxifen, toremifene, fluoxymesterol andraloxifene), antiandrogens (bicalutamide, nilutamide, and flutamide),aromatase inhibitors (e.g., aminoglutethimide, anastrozole andtetrazole), ketoconazole, goserelin acetate, leuprolide, megestrolacetate and mifepristone. Combination therapy including a compounddisclosed herein and a hormonal agent is expected to have therapeuticsynergistic effects on cancer and reduce sides affects associated withthese chemotherapeutic agents. Plant-derived agents useful incombination with the compounds disclosed herein include, but are notlimited to, vinca alkaloids (e.g., vincristine, vinblastine, vindesine,vinzolidine and vinorelbine), podophyllotoxins (e.g., etoposide (VP-16)and teniposide (VM-26)), taxanes (e.g., paclitaxel and docetaxel). Theseplant-derived agents generally act as antimitotic agents that bind totubulin and inhibit mitosis. Podophyllotoxins such as etoposide arebelieved to interfere with DNA synthesis by interacting withtopoisomerase II, leading to DNA strand scission. Combination therapyincluding a compound disclosed herein and a plant-derived agent isexpected to have therapeutic synergistic effects on cancer and reducesides affects associated with these chemotherapeutic agents

In any case, regardless of the disease, disorder or condition beingtreated, the overall benefit experienced by the patient is simply beadditive of the two therapeutic agents or the patient experiences asynergistic benefit.

In certain embodiments, different therapeutically-effective dosages ofthe compounds disclosed herein will be utilized in formulatingpharmaceutical composition and/or in treatment regimens when thecompounds disclosed herein are administered in combination with one ormore additional agent, such as an additional therapeutically effectivedrug, an adjuvant or the like. Therapeutically-effective dosages ofdrugs and other agents for use in combination treatment regimens isoptionally determined by means similar to those set forth hereinabovefor the actives themselves. Furthermore, the methods ofprevention/treatment described herein encompasses the use of metronomicdosing, i.e., providing more frequent, lower doses in order to minimizetoxic side effects. In some embodiments, a combination treatment regimenencompasses treatment regimens in which administration of a compounddescribed herein, or a pharmaceutically acceptable salt thereof, isinitiated prior to, during, or after treatment with a second agentdescribed herein, and continues until any time during treatment with thesecond agent or after termination of treatment with the second agent. Italso includes treatments in which a compound described herein, or apharmaceutically acceptable salt thereof, and the second agent beingused in combination are administered simultaneously or at differenttimes and/or at decreasing or increasing intervals during the treatmentperiod. Combination treatment further includes periodic treatments thatstart and stop at various times to assist with the clinical managementof the patient.

It is understood that the dosage regimen to treat, prevent, orameliorate the condition(s) for which relief is sought, is modified inaccordance with a variety of factors (e.g. the disease, disorder orcondition from which the subject suffers; the age, weight, sex, diet,and medical condition of the subject). Thus, in some instances, thedosage regimen actually employed varies and, in some embodiments,deviates from the dosage regimens set forth herein.

For combination therapies described herein, dosages of theco-administered compounds vary depending on the type of co-drugemployed, on the specific drug employed, on the disease or conditionbeing treated and so forth. In additional embodiments, whenco-administered with one or more other therapeutic agents, the compoundprovided herein is administered either simultaneously with the one ormore other therapeutic agents, or sequentially.

In combination therapies, the multiple therapeutic agents (one of whichis one of the compounds described herein) are administered in any orderor even simultaneously. If administration is simultaneous, the multipletherapeutic agents are, by way of example only, provided in a single,unified form, or in multiple forms (e.g., as a single pill or as twoseparate pills).

The compounds described herein, or a pharmaceutically acceptable saltthereof, as well as combination therapies, are administered before,during or after the occurrence of a disease or condition, and the timingof administering the composition containing a compound varies. Thus, inone embodiment, the compounds described herein are used as aprophylactic and are administered continuously to subjects with apropensity to develop conditions or diseases in order to prevent theoccurrence of the disease or condition. In another embodiment, thecompounds and compositions are administered to a subject during or assoon as possible after the onset of the symptoms. In specificembodiments, a compound described herein is administered as soon as ispracticable after the onset of a disease or condition is detected orsuspected, and for a length of time necessary for the treatment of thedisease. In some embodiments, the length required for treatment varies,and the treatment length is adjusted to suit the specific needs of eachsubject. For example, in specific embodiments, a compound describedherein or a formulation containing the compound is administered for atleast 2 weeks, about 1 month to about 5 years.

In some embodiments, a compound described herein, or a pharmaceuticallyacceptable salt thereof, is administered in combination withchemotherapy, hormone blocking therapy, radiation therapy, monoclonalantibodies, or combinations thereof.

Chemotherapy includes the use of one or more anti-cancer agents.

EXAMPLES

The following examples are provided for illustrative purposes only andnot to limit the scope of the claims provided herein.

Having now generally described the invention, the same will be morereadily understood through reference to the following examples which areprovided by way of illustration, and are not intended to be limiting ofthe present invention, unless specified.

Example 1: Synthesis of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylatesodium salt (Compound 1A)

Step 1: Synthesis of methyl2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid and Ethyl2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid

To a suspension of 2,6-dichloronicotinaldehyde (3.55 g, 20.2 mmol) andmethyl benzimidazole-2-acetate (3.82 g, 20.1 mmol, 1 eq.) in ethanol (50mL) was added N-methylhomopiperazine (4.66 ml, 37.46 mmol) and themixture was stirred at room temperature for 20 min. The reaction mixturewas heated to reflux for 6 h. The mixture was then cooled to −20° C. andthe precipitate collected by filtration to give a first crop of product(2.61 g). The filtrate was concentrated and the residue triturated withethyl acetate (50 mL) to give a second crop (1.58 g). The filtrate wasconcentrated and the residue triturated with a mixture of methanol:water(20 mL, 1:1 v/v) to give a third (748 mg). The combined crops containeda mixture of the title compounds (4.94 g, 12.4 mmol, 61%) and were usedwithout further purification. LCMS: 98% (8.8:1 OMe:OEt), Rt 1.434 min,ESMS m/z 390.1 (M+H)⁺ (Me ester), Rt 1.575 min, ESMS m/z 404.1 (M+H)⁺(Et ester).

Step 2: Synthesis of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid Sodium Salt (1A)

A suspension of methyl and ethyl2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid (combined products from the previous step, 2.77 g, 7.1 mmol) andsodium hydroxide (284 mg, 14.2 mmol, 2 eq.) in a mixture of water and1,4-dioxane (30 mL, 1:1 v/v) was heated to 60° C. for 15 min. Themixture was cooled to room temperature and the precipitate collected.The solid was washed with acetonitrile (2×10 mL) and ether (2×10 mL) andair dried to yield the title compound (2.61 g, 6.6 mmol, 92%) as yellowsolid. LCMS: 91%, Rt: 1.386 min, ESMS m/z 376.0 (M+H)⁺.

Example 2: Synthesis of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid isopropylamide (Compound 2A)

A mixture of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid sodium salt (1A, 127 mg, 0.32 mmol), isopropylamine (28 mg, 0.48mmol, 1.5 eq.),1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxide hexafluorophosphate (HATU, 182 mg, 0.48 mmol, 1.5 eq.) anddiisopropylethylamine (62 mg, 0.48 mmol, 1.5 eq.) inN,N-dimethylformamide (6.4 mL) was stirred at room temperature for 1 h.

The reaction mixture was poured into water (20 mL) and extracted withchloroform (3×10 mL). The combined organic layers were washed withsaturated aqueous ammonium chloride (2×2 mL), water (10 mL) and brine(10 mL), dried over magnesium sulfate and evaporated. The residue wastriturated with hexane (2 mL) to give the title compound (45 mg, 0.108mmol, 34%) as a yellow crystalline solid.

LCMS: 92%, Rt: 1.886 min, ESMS m/z 417.2 (M+H)⁺. ¹H NMR (300 MHz, CDCl₃)δ 10.51 (d, J=7.4 Hz, 1H), 8.99 (d, J=8.0 Hz, 1H), 8.63 (s, 1H), 7.98(d, J=9.0 Hz, 1H), 7.95 (d, J=7.8 Hz, 1H), 7.57 (t, J=7.2 Hz, 1H), 7.46(t, J=7.2 Hz, 1H), 6.72 (d, J=8.9 Hz, 1H), 4.43 (sex, J=6.7 Hz, 1H),4.25-4.00 (m, 2H), 4.00-3.75 (m, 2H), 3.05-2.83 (m, 2H), 2.74-5.59 (m,2H), 2.45 (s, 3H), 2.30-2.10 (m, 2H), 1.44 (d, J=6.6 Hz, 6H).

Example 3: Synthesis of2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid (1-cyclopropylethyl)-amide (Compound 2B)

A mixture of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid sodium salt (1A, 127 mg, 0.32 mmol), 1-cyclopropylethylamine (41mg, 0.48 mmol, 1.5 eq.),1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxide hexafluorophosphate (HATU, 182 mg, 0.48 mmol, 1.5 eq.) anddiisopropylethylamine (62 mg, 0.48 mmol, 1.5 eq.) inN,N-dimethylformamide (6.4 mL) was stirred at room temperature for 1 h.

The reaction mixture was poured into water (20 mL) and extracted withchloroform (3×10 mL). The combined organic layers were washed withsaturated aqueous ammonium chloride (2×2 mL), water (10 mL) and brine(10 mL), dried over magnesium sulfate and evaporated. The residue waspurified by silica gel column chromatography eluting withchloroform:methanol (100:0→90:10) to give the title compound (10 mg,0.023 mmol, 7%) as a pale yellow crystalline solid.

LCMS: 97%, Rt: 1.926 min, ESMS m/z 443.1 (M+H)⁺. ¹H NMR (300 MHz, CDCl₃)δ 10.59 (d, J=8.0 Hz, 1H), 8.87 (d, J=8.2 Hz, 1H), 8.61 (s, 1H), 7.96(d, J=8.6 Hz, 1H), 7.94 (d, J=8.6 Hz, 1H), 7.58 (t, J=7.5 Hz, 1H), 7.44(t, J=7.5 Hz, 1H), 6.71 (d, J=8.8 Hz, 1H), 4.40-4.09 (m, 2H), 3.99-3.72(m, 3H), 3.31-3.04 (m, 2H), 3.04-2.78 (m, 2H), 2.61 (s, 3H), 2.52-2.28(m, 2H), 1.46 (d, J=6.6 Hz, 3H), 1.23-1.06 (m, 1H), 0.65-0.44 (m, 3H),0.41-0.28 (m, 1H).

Example 4: Synthesis of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid cyclopropylamide (Compound 2C)

A mixture of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid sodium salt (1A, 127 mg, 0.32 mmol), cyclopropylamine (27 mg, 0.48mmol, 1.5 eq.),1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxide hexafluorophosphate (HATU, 182 mg, 0.48 mmol, 1.5 eq.) anddiisopropylethylamine (62 mg, 0.48 mmol, 1.5 eq.) inN,N-dimethylformamide (6.4 mL) was stirred at room temperature for 1 h.

The reaction mixture was poured into water (20 mL) and extracted withchloroform (3×10 mL). The combined organic layers were washed withsaturated aqueous ammonium chloride (4×2 mL), water (10 mL) and brine(10 mL), dried over magnesium sulfate and evaporated to give the titlecompound (25 mg, 0.060 mmol, 19%) as a yellow crystalline solid. LCMS:95%, Rt: 1.747 min, ESMS m/z 415.3 (M+H)⁺. ¹H NMR (300 MHz, CDCl₃) δ10.58 (d, J=3.8 Hz, 1H), 8.88 (d, J=8.1 Hz, 1H), 8.60 (s, 1H), 7.96 (d,J=9.3 Hz, 1H), 7.93 (d, J=9.3 Hz, 1H), 7.55 (t, J=7.6 Hz, 1H), 7.44 (t,J=7.7 Hz, 1H), 6.69 (d, J=8.9 Hz, 1H), 4.32-4.02 (m, 2H), 3.99-3.75 (m,2H), 3.18-2.94 (m, 3H), 2.91-2.72 (m, 2H), 2.54 (s, 3H), 2.42-2.23 (m,2H), 1.04-0.91 (m, 2H), 0.88-0.74 (m, 2H).

Example 5: Synthesis of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid (tetrahydropyran-4-yl)-amide (Compound 2D)

A mixture of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid sodium salt (1A, 127 mg, 0.32 mmol), 4-aminotetrahydropyran (48 mg,0.48 mmol, 1.5 eq.),1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxide hexafluorophosphate (HATU, 182 mg, 0.48 mmol, 1.5 eq.) anddiisopropylethylamine (62 mg, 0.48 mmol, 1.5 eq.) inN,N-dimethylformamide (6.4 mL) was stirred at room temperature for 1 h.

The reaction mixture was poured into water (20 mL) and extracted withchloroform (3×10 mL). The combined organic layers were washed withsaturated aqueous ammonium chloride (2×2 mL), water (10 mL) and brine(10 mL), dried over magnesium sulfate and evaporated. The residue wastriturated with 1:1 hexane:ether (2 mL) to give the title compound (17mg, 0.037 mmol, 12%) as a yellow crystalline solid. LCMS: 95%, Rt: 1.780min, ESMS m/z 459.1 (M+H)⁺. ¹H NMR (300 MHz, CDCl₃-d) δ 10.76 (d, J=7.6Hz, 1H), 8.94 (d, J=8.2 Hz, 1H), 8.61 (s, 1H), 7.97 (d, J=9.6 Hz, 1H),7.93 (d, J=9.3 Hz, 1H), 7.56 (t, J=7.6 Hz, 1H), 7.45 (t, J=7.7 Hz, 1H),6.71 (d, J=8.9 Hz, 1H), 4.48-4.28 (m, 1H), 4.21-3.99 (m, 4H), 3.99-3.81(m, 2H), 3.75-3.57 (m, 2H), 3.10-2.88 (m, 2H), 2.82-2.63 (m, 2H), 2.48(s, 3H), 2.35-2.20 (m, 2H), 2.17-2.07 (m, 2H), 1.96-1.74 (m, 2H).

Example 6: Synthesis of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid (5-methylpyrazin-2-ylmethyl)-amide (Compound 2E)

A mixture of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid sodium salt (1A, 127 mg, 0.32 mmol),2-(aminomethyl)-5-methylpyrazine (59 mg, 0.48 mmol, 1.5 eq.),1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxide hexafluorophosphate (HATU, 182 mg, 0.48 mmol, 1.5 eq.) anddiisopropylethylamine (62 mg, 0.48 mmol, 1.5 eq.) inN,N-dimethylformamide (6.4 mL) was stirred at room temperature for 16 h.

The reaction mixture was poured into water (20 mL) and the precipitatecollected by filtration to give the title compound (23 mg, 0.048 mmol,15%) as a yellow crystalline solid. LCMS: 96%, Rt: 1.712 min, ESMS m/z481.3 (M+H)⁺. ¹H NMR (300 MHz, CDCl₃) δ 11.34-11.16 (m, 1H), 8.97 (d,J=8.1 Hz, 1H), 8.68 (s, 1H), 8.63 (s, 1H), 8.47 (s, 1H), 7.96 (d, J=8.6Hz, 1H), 7.93 (d, J=8.6 Hz, 1H), 7.55 (t, J=7.6 Hz, 1H), 7.44 (t, J=7.7Hz, 1H), 6.72 (d, J=9.0 Hz, 1H), 4.98 (d, J=5.6 Hz, 2H), 4.23-3.73 (m,4H), 3.00-2.80 (m, 2H), 2.72-2.61 (m, 2H), 2.59 (s, 3H), 2.43 (s, 3H),2.25-2.06 (m, 2H).

Example 7: Synthesis of2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid (3-methyloxetan-3-ylmethyl)-amide (Compound 2F)

A mixture of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid sodium salt (1A, 127 mg, 0.32 mmol),(3-methyloxetan-3-yl)methanamine (48 mg, 0.48 mmol, 1.5 eq.),1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxide hexafluorophosphate (HATU, 182 mg, 0.48 mmol, 1.5 eq.) anddiisopropylethylamine (62 mg, 0.48 mmol, 1.5 eq.) inN,N-dimethylformamide (6.4 mL) was stirred at room temperature for 16 h.

The reaction mixture was poured into water (20 mL) and extracted withchloroform (3×10 mL). The combined organic layers were washed withsaturated aqueous ammonium chloride (2×2 mL), water (10 mL) and brine(10 mL), dried over magnesium sulfate and evaporated to give the titlecompound (6 mg, 0.013 mmol, 4%) as a yellow crystalline solid. LCMS 99%,Rt: 1.836 min, ESMS m/z 459.1 (M+H)⁺. ¹H NMR (300 MHz, CDCl₃-d) δ11.08-10.85 (m, 1H), 8.99 (d, J=8.1 Hz, 1H), 8.63 (s, 1H), 7.98 (d,J=8.9 Hz, 1H), 7.90 (d, J=8.1 Hz, 1H), 7.55 (t, J=7.6 Hz, 1H), 7.45 (t,J=7.7 Hz, 1H), 6.74 (d, J=8.9 Hz, 1H), 4.74 (d, J=6.0 Hz, 2H), 4.47 (d,J=5.9 Hz, 2H), 4.33-3.65 (m, 4H), 3.86 (d, J=6.0 Hz, 2H), 3.00-2.83 (m,2H), 2.74-2.56 (m, 2H), 2.43 (s, 3H), 2.28-2.11 (m, 2H), 1.52 (s, 3H).

Example 8: Synthesis of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid methylamide (Compound 2G)

A mixture of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid sodium salt (1A, 99 mg, 0.25 mmol), methylamine hydrochloride (25mg, 0.38 mmol, 1.5 eq.),1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxide hexafluorophosphate (HATU, 144 mg, 0.38 mmol, 1.5 eq.) anddiisopropylethylamine (49 mg, 0.38 mmol, 1.5 eq.) inN,N-dimethylformamide (5.0 mL) was stirred at room temperature for 1 h.

The reaction mixture was poured into water (8 mL) and cooled to 4° C.for 16 h. The precipitate was collected and washed with 4:1 water:N,N-dimethylformamide (1 mL) and water (1 mL). The solid was suspendedin water (200 μL) and 20% aqueous sodium carbonate (200 μL) was added.The mixture was stirred at room temperature for 1 h, the solid wascollected and washed with water (3×1 mL) to give the title compound (16mg, 0.041 mmol, 16%) as a yellow crystalline solid. LCMS: 100%, Rt:1.678 min, ESMS m/z 389.1 (M+H)⁺. ¹H NMR (300 MHz, CDCl₃-d) δ10.56-10.34 (m, 1H), 8.99 (d, J=8.1 Hz, 1H), 8.64 (s, 1H), 7.96 (d,J=9.0 Hz, 2H), 7.95 (d, J=7.8 Hz, 1H), 7.57 (t, J=7.8 Hz, 1H), 7.47 (t,J=7.8 Hz, 1H), 6.71 (d, J=9.0 Hz, 1H), 4.41-3.69 (m, 4H), 3.21 (d, J=4.8Hz, 3H), 3.00-2.81 (m, 2H), 2.74-2.58 (m, 2H), 2.44 (s, 3H), 2.27-2.07(m, 2H).

Example 9: Synthesis of2-((3S,5R)-3,5-dimethylpiperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile(Compound 3A)

To a suspension of 2,6-dichloronicotinaldehyde (100 mg, 0.568 mmol) andmethyl benzimidazole-2-acetate (89.5 mg, 0.568 mmol, 1 eq.) in ethanol(1.5 mL) was added cis-2,6-dimethylpiperazine (120.8 mg, 1.058 mmol) andthe mixture was heated to 75° C. for 24 hours.

The mixture was then cooled to room temperature and the precipitatecollected by filtration, washed with ethyl acetate and small amount ofmethanol, and dried under vacuum to give orange solid (42 mg).

Example 10: Synthesis of2-(4-ethylpiperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile(Compound 3B)

To a suspension of 2,6-dichloronicotinaldehyde (100 mg, 0.568 mmol) andmethyl benzimidazole-2-acetate (89.5 mg, 0.568 mmol, 1 eq.) in ethanol(1.5 mL) was added 1-ethylpiperazine (120.8 mg, 1.058 mmol) and themixture was heated to 75° C. for 24 hours.

The mixture was then cooled to room temperature and the precipitatecollected by filtration, washed with ethyl acetate and small amount ofmethanol and dried under vacuum to give orange solid (45 mg).

Example 11: Synthesis of Ethyl 1H-benzimidazol-2-ylacetate

A mixture of SM1 (40 g, 370 mmol) and SM2 (73 g, 373 mmol) in ethanol(200 ml) was stirred at 90° C. overnight. The reaction was cooled toroom temperature and the solvent was removed in vacuo. Water (300 ml)and DCM (500 ml) were added to the residue. The organic layer wasseparated, dried over Na₂SO₄ and the solvent removed to give Ethyl1H-benzimidazol-2-ylacetate (40 g, 53% yield) as a pale yellow solid.

Example 12: Synthesis of Ethyl2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid

To a suspension of SM1 (25.9 g, 148 mmol) and SM2 (30 g, 147 mmol) inethanol (400 mL) was added SM3 (33.5 g, 394 mmol) and the mixture wasstirred at room temperature for 20 min. The reaction mixture was heatedto reflux for 6 h. The mixture was then cooled to 20° C. and theprecipitate collected by filtration to give Ethyl2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid (40 g, 68% yield) as a pale yellow solid. LCMS

Example 13: Synthesis of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid

A suspension of Ethyl2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid (1.2 g, 3.0 mmol) and sodium hydroxide (238 mg, 6.0 mmol) in amixture of water and 1,4-dioxane (10 mL, 1:1 v/v) was heated to 60° C.for 30 min. The mixture was concentrated under reduced pressure. Themixture was acidified to pH 6 by addition of 1 M hydrochloric acid andevaporated to give the product (1.0 g, yield=89%), as a yellowcrystalline solid.

Example 14: Synthesis of Compound 4A

A mixture of2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid (150 mg, 0.4 mmol), thiazol-2-amine (60 mg, 0.6 mmol), HATU (228mg, 0.6 mmol) and diisopropylethylamine (158 mg, 1.2 mmol) inN,N-dimethylformamide (5 mL) was stirred at room temperature over night.The yellow precipitate was collected and washed with ethyl alcohol anddiethyl ether to give the crude product.

This material was dissolved in 2 M aqueous sodium hydroxide (20 mL) andextracted with dichloromethane (100 mL, then 20 mL). The combinedorganic layers were dried over magnesium sulfate and evaporated. Theresidue was triturated with diethyl ether (30 mL) and washed withdiethyl ether to give the title compound (30 mg) as a yellow crystallinesolid.

Example 15: Synthesis of Compound 4B

Compound 4B was synthesized according to the procedure of 4A.

Example 16: Synthesis of Compound 4C

Compound 4C was synthesized according to the procedure of 4A.

Example 17: Synthesis of Compound 5A

To a suspension of SM1 (4.32 g, 25.0 mmol) and SM2 (5.0 g, 25.0 mmol) inethanol (100 mL) was added SM3 (4.9 g, 50.0 mmol) and the mixture wasstirred at room temperature for 20 min. The reaction mixture was heatedto reflux for 6 h. The mixture was then cooled to 20° C. and theprecipitate collected by filtration, washed with EtOH, Et₂O and dried togive compound 5A (5.0 g, yield=50%, Lot#: MC13021-014-03) as a paleyellow solid. LCMS: m/z 390 (M+H)⁺.

Example 18: Synthesis of Compound 5B

A suspension of compound 5A (100 mg, 0.257 mmol) and 85% NH₂NH₂.H₂O (300mg, 5.0 mmol) in EtOH (20 ml) was heated to reflux for 6 h. The mixturewas then cooled to 20° C. and the precipitate collected by filtration,washed with EtOH and dried to give compound 5B (40 mg, yield=41%) as apale yellow solid. LCMS: m/z 376 (M+H)⁺.

Example 19: Synthesis of Compound 5C

A suspension of compound 5A (100 mg, 0.257 mmol) in CH₃NH₂/EtOH (2M, 20ml, 40 mmol) was heated to reflux for 6 h. The mixture was then cooledto 20° C. and the precipitate collected by filtration, washed with EtOHand dried. The yellow solid was dissolved in DCM (100 ml) and it waswashed with water (3 times), brine, dried and concentrated to givecompound 5C (20 mg, yield=21%) as a pale yellow solid.

Example 20: Synthesis of Compound 5D

A suspension of compound 5A (100 mg, 0.257 mmol) and propan-2-amine (295mg, 5.0 mmol) in EtOH (20 ml) was heated to reflux for 6 h. The mixturewas then cooled to 20° C. and the precipitate collected by filtration,washed with EtOH and dried to give compound 5D (20 mg, yield=19%) as apale yellow solid. LCMS: m/z 403.3 (M+H)⁺.

Example 21: Synthesis of Compound 2G

A suspension of Ethyl2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triazabenzo[c]fluorene-6-carboxylicacid (40 g, 99 mmol) in CH₃NH₂/EtOH (2M, 400 ml, 800 mmol) was heated toreflux for 6 h. The mixture was then cooled to 20° C. and theprecipitate was collected by filtration to give crude 2G as a paleyellow solid (35 g, 86% yield). The crude product was dissolved in DCM(500 ml) and washed with water (3 times), brine, dried and concentratedto give 2G (30.2 g, yield=78%) as a pale yellow solid.

1H NMR (400 MHz, DMSO-d6) δ 10.18 (1H, d, J=4.8 Hz), 8.91 (1H, d, J=6.0Hz), 8.58 (1H, s), 8.25 (1H, d, J=8.8 Hz), 7.91 (1H, d, J=8.0 Hz), 7.55(1H, t, J=7.6 Hz), 7.47 (1H, t, J=7.6 Hz), 6.99 (1H, d, J=88 Hz),4.06-3.95 (4H, m), 3.03 (3H, d, J=4.8 Hz), 2.79-2.76 (2H, m), 2.50 (2H,m), 2.28 (3H, s), 2.02 (2H, br s). LC-MS: Rt=9.160 min, 389.5[M+H]⁺.

Example 22: Synthesis of2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile(Compound 3C)

To a solution of compound SM1 (100 mg, 0.568 mmol) and compound SM2 (90mg, 0.568 mmol) in 2-Methoxyethanol (10 mL) was addedN-Methylhomopiperazine (130 mg, 1.136 mmol) and the mixture was stirredat room temperature for 20 min. The reaction mixture was heated toreflux for 16 h. The mixture was then cooled to 20° C. and theprecipitate collected by filtration, washed with EtOH, Et₂O, dried andpurified by Pre-HPLC to give 3C (15 mg) as a pale yellow solid. LCMS:m/z 357 (M+H)+.

Example 23: Synthesis of2-[1,4]Diazepan-1-yl-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile(Compound 3D)

Compound 3D was synthesized as pale yellow solid (20 mg) according tothe procedure of compound 3C. LCMS: m/z 342.9 (M+H)+.

Example 24: Synthesis of2-(2-Morpholin-4-yl-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile(Compound 3E)

Compound 3E was synthesized as pale yellow solid (16 mg) according tothe procedure of compound 3C. LCMS: m/z 373 (M+H)+.

Example 25: Synthesis of2-(3-Dimethylamino-pyrrolidin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile(Compound 3F)

Compound 3F was synthesized as pale yellow solid (15 mg) according tothe procedure of compound 3C. LCMS: m/z 357 (M+H)+.

Example 26: Synthesis of2-(2-Imidazol-1-yl-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile(Compound 3G)

Compound 3G was synthesized as pale yellow solid (12 mg) according tothe procedure of compound 3C. LCMS: m/z 354 (M+H)+.

Example 27: Synthesis of2-(4-Methyl-piperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile(3H)

Compound 3H was synthesized as pale yellow solid (30 mg) according tothe procedure of compound 3C. LCMS: m/z 342.9 (M+H)+.

Example 28: Synthesis of2-(2-Methoxy-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile(Compound 3I)

Compound 3I was synthesized as pale yellow solid (15 mg) according tothe procedure of compound 3C. LCMS: m/z 318 (M+H)+.

Example 29: Synthesis of2-(1-Methyl-piperidin-4-ylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile(Compound 3J)

Compound 3J was synthesized as pale yellow solid (7 mg) according to theprocedure of compound 3C. LCMS: m/z 357 (M+H)+.

Example 30: Synthesis of2-(2-Dimethylamino-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carbonitrile(Compound 3K)

Compound 3K was synthesized as pale yellow solid (7 mg) according to theprocedure of compound 3C. LCMS: m/z 331 (M+H)+.

Example 31: Synthesis of2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid N′-methyl-hydrazide (Compound 4D)

To a solution of2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid ethyl ester (100 mg, 0.25 mmol) in ethanol (10 mL) was addedmethylhydrazine (288 mg, 40% aqueous solution, 2.5 mmol) and the mixturewas stirred at room temperature for 20 min. The reaction mixture washeated to reflux for 12 hrs. The mixture was then cooled to 20° C.,concentrated and washed with water and methanol, dried to give compound4D (20 mg, yield=20%) as yellow solid. LCMS: m/z 404 (M+H)+.

Example 32: Synthesis of2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid (Compound 1A)

A suspension of2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid ethyl ester (1.2 g, 3.0 mmol) and 2N NaOH (aq., 3 mL, 6.0 mmol) ina mixture of water and EtOH (10 mL, 1:1 v/v) was heated to 60° C. for 30min. The mixture was concentrated under reduced pressure. The mixturewas acidified to pH 6 by addition of 2 N HCl and stirred at r.t. for 30min, the mixture was filtered and washed with water and ethanol, driedto give the desired product as a yellow solid (1.0 g, 89% yield). LCMS:m/z 376 (M+H)+.

Example 33: Synthesis of2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid methyl-d₃-amide (Compound 4E)

Example 34: Synthesis of2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid methoxy-amide (Compound 4F)

Compound 4E was synthesized as yellow solid (40 mg) according to theprocedure of compound 4A. LCMS: m/z 392.6 (M+H)+.

Compound 4F was synthesized as yellow solid (14 mg) according to theprocedure of compound 4A. LCMS: m/z 305 (M+H)+.

Example 35: Synthesis of1,1-Dimethyl-4-(6-methylcarbamoyl-1,7,11b-triaza-benzo[c]fluoren-2-yl)-[1,4]diazepan-1-ium;iodide (Compound 6A)

To a mixture of compound 2G (25 mg, 0.064 mmol), and K₂CO₃ (18 mg, 0.128mmol) in N,N-dimethylformamide (2 mL) was added MeI (11 mg, 0.076 mmol)and stirred at 50° C. for 1 h. After cooling, the mixture was filteredand the filtrate was purified by Pre-HPLC to give the title compound 6A(16 mg, 47% yield) as yellow solid. LCMS: m/z 403.7 (M+H)+.

Example 36: Synthesis of2-(4-Methyl-piperazin-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid cyclopropylamide (Compound 5A)

Compound 5E was synthesized as yellow solid (25 mg) according to theprocedure of compound 4D. LCMS: m/z 401 (M+H)+.

Example 37: Synthesis of3-Fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic acid ethyl ester(Compound 7A)

To a solution of compound SM1 (0.8 g, 5 mmol) and compound SM2 (1.02 g,5 mmol) in ethanol (50 mL) was added piperidine (1.28 g, 15 mmol) andthe mixture was stirred at room temperature for 20 min. The reactionmixture was heated to reflux for 16 h. After cooling, the mixture wasfiltered and washed with ethanol, dried to get compound 7A as yellowsolid (0.82 g, 79% yield). LCMS: m/z 309.9 (M+H)+.

Example 38: Synthesis of3-Fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic acid (Compound 7B)

Compound 7B was synthesized as yellow solid (320 mg) according to theprocedure of compound 1A. LCMS: m/z 281.9 (M+H)+.

Example 39: Synthesis of(3-Fluoro-1,7,11b-triaza-benzo[c]fluoren-6-yl)-(4-methyl-[1,4]diazepan-1-yl)-methanone(Compound 7C)

Compound 7C was synthesized as yellow solid (20 mg) according to theprocedure of compound 4A. LCMS: m/z 378.5 (M+H)+.

Example 40: Synthesis of[1,4]Diazepan-1-yl-(3-fluoro-1,7,11b-triaza-benzo[c]fluoren-6-yl)-methanone(Compound 7D)

A mixture of 7B (45 mg, 0.16 mmol), N-Boc-homopiperazine (64 mg, 0.32mmol), HOBt (43.2 mg, 0.32 mmol), DIEA (103.8 mg, 0.81 mmol) andEDCI.HCl (62 mg, 0.32 mmol) in N,N-dimethylformamide (5 mL) was stirredat room temperature overnight. This material was dissolved in water (20ml) and extracted with 10% MeOH in DCM (20 mL). The organic layers weredried over magnesium sulfate and evaporated. The residue was dissolvedin MeOH (5 mL) and 4N HCl (0.5 mL) was added and the mixture was stirredat r.t. for 4 h. The mixture was adjusted to pH˜8 by sat. NaHCO3,filtered, washed with ether to give compound 7D as pale yellow solid (40mg, 68% yield). LCMS: m/z 364.5 (M+H)+.

Example 41: Synthesis of3-Fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic acid(1-methyl-piperidin-4-yl)-amide (Compound 7E)

Compound 7E was synthesized as yellow solid (32 mg) according to theprocedure of compound 4A. LCMS: m/z 378.5 (M+H)+.

Example 42: Synthesis of3-Fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic acid(2-dimethylamino-ethyl)-amide (Compound 7F)

A solution of 7B (50 mg, 0.16 mmol) and N1,N1-dimethylethane-1,2-diamine(295 mg, 5.0 mmol) in EtOH (10 ml) was heated to reflux for 16 h. Themixture was then cooled to r.t., filtered and washed by ether, dried togive compound 7F (20 mg, 35% yield) as a pale yellow solid. LCMS: m/z352.4 (M+H)+.

Example 43: Synthesis of3-Fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic acid(2-morpholin-4-yl-ethyl)-amide (Compound 7G)

Compound 7G was synthesized as yellow solid (24 mg) according to theprocedure of compound 7F. LCMS: m/z 394 (M+H)+.

Example 44: Synthesis of3-Fluoro-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic acid(2-pyrrolidin-1-yl-ethyl)-amide (Compound 7I)

Compound 7I was synthesized as yellow solid (21 mg) according to theprocedure of compound 7F. LCMS: m/z 378 (M+H)+.

Example 45: Synthesis of4-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid ethyl ester (Compound 8A)

To a solution of SM1 (176 mg, 1 mmol) and SM2 (204 mg, 1 mmol) in2-Methoxyethanol (10 mL) was added N-Methylhomopiperazine (342 mg, 3mmol) and the mixture was stirred at room temperature for 20 min. Thereaction mixture was heated to reflux for 16 h. The crude product waspurification by gel silica column (DCM/MeOH=50:1) to get compound 8A asbrown oil (320 mg, 79% yield). LCMS: m/z 404 (M+H)+.

Example 46: Synthesis of4-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid methylamide (Compound 8B)

A suspension of compound 8A (80 mg, 0.198 mmol) in CH₃NH₂/EtOH (2M, 4ml, 800 mmol) was heated to reflux for 16 h. After cooling, the mixturewas filtered and washed with ethanol, dried to give compound 8B as apale yellow solid (40 mg, 52% yield). LCMS: m/z 389 (M+H)+.

Example 47: Synthesis of Compound 8C

Compound 8C was synthesized as yellow solid (360 mg) according to theprocedure of compound 8A. LCMS: m/z 404 (M+H)+.

Example 48: Synthesis of Compound 8D

Compound 8C was synthesized as yellow solid (40 mg) according to theprocedure of compound 8B. LCMS: m/z 389 (M+H)+.

Example 49: Synthesis of4-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid (Compound 8E)

Compound 8E was synthesized as pale yellow solid (140 mg) according tothe procedure of compound 1A. LCMS: m/z 376 (M+H)+.

Example 50: Synthesis of4-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid isopropylamide (Compound 8F)

Compound 8F was synthesized as yellow solid (10 mg) according to theprocedure of compound 4A. LCMS: m/z 417 (M+H)+.

Example 51: Synthesis of4-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid methoxy-amide (Compound 8G)

Compound 8G was synthesized as yellow solid (16 mg) according to theprocedure of compound 4A. LCMS: m/z 405 (M+H)+.

Example 52: Synthesis of Compound 8H

Compound 8H was synthesized as pale yellow solid (180 mg) according tothe procedure of compound 1A. LCMS: m/z 376 (M+H)+.

Example 53: Synthesis of Compound 8I

Compound 8I was synthesized as yellow solid (10 mg) according to theprocedure of compound 4A. LCMS: m/z 417 (M+H)+.

Example 54: Synthesis of Compound 8J

Compound 8J was synthesized as yellow solid (8 mg) according to theprocedure of compound 4A. LCMS: m/z 415 (M+H)+.

Example 55: Synthesis of Compound 8K

Compound 8K was synthesized as yellow solid (11 mg) according to theprocedure of compound 4A. LCMS: m/z 405 (M+H)+.

Example 56: Synthesis of2-(4-tert-Butoxycarbonyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid ethyl ester (Compound 9A)

To a solution of SM1 (176 mg, 1 mmol) and SM2 (204 mg, 1 mmol) in2-Methoxyethanol (10 mL) was added N-Bochomopiperazine (600 mg, 3 mmol)and the mixture was stirred at room temperature for 20 min. The reactionmixture was heated to reflux for 16 h. The crude product waspurification by gel silica column (DCM/MeOH=50:1) to get compound 9A asbrown oil (490 mg, 50% yield). LCMS: m/z 490 (M+H)+.

Example 57: Synthesis of2-[1,4]Diazepan-1-yl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic acidisopropylamide (Compound 9B)

A solution of compound 9A (100 mg, 0.20 mmol) and propan-2-amine (295mg, 5.0 mmol) in EtOH (10 ml) was heated to reflux for 6 h. The mixturewas then cooled to 20° C. and concentrated. To the residue was added 2NHCl (1 ml, 2.0 mmol) in MeOH (5 mL) and the mixture is stirred at r.t,overnight. The mixture was concentrated and purified by Pre-HPLC to givecompound 9B (12 mg, yield=12%) as a pale yellow solid. LCMS: m/z 403(M+H)+.

Example 58: Synthesis of2-[1,4]Diazepan-1-yl-1,7,11b-triaza-benzo[c]fluorene-6-carboxylic acidmethylamide (Compound 9C)

Compound 9C was synthesized as yellow solid (20 mg) according to theprocedure of compound 4A. LCMS: m/z 375 (M+H)+.

Example 59: Synthesis ofN-Hydroxy-2-(4-methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxamidine(Compound 10A)

To a solution of compound 3C (356 mg, 1 mmol) and NH₂OH.HCl (83 mg, 1.2mmol) in methanol (10 mL) was added 1N NaHCO₃ (1.2 mL, 1.2 mmol) and themixture was stirred at refluxing for 6 h. After cooling, the mixture wasfiltered and washed with methanol to give compound 10A (210 mg, 53%yield) as yellow solid. LCMS: m/z 390 (M+H)+.

Example 60: Synthesis of3-[2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluoren-6-yl]-4H-[1,2,4]oxadiazol-5-one(Compound 10B)

A mixture of compound 10A (78 mg, 0.2 mmol) and CDI (83 mg, 0.3 mmol) inCH₃CN (5 mL) was stirred at refluxing for 6 h. The mixture wasconcentrated and purified by pre-HPLC to give compound 10B (16 mg, 19%yield) as yellow solid. LCMS: m/z 416 (M+H)+.

Example 61: Synthesis of2-(4-Methyl-[1,4]diazepan-1-yl)-6-(1H-tetrazol-5-yl)-1,7,11b-triaza-benzo[c]fluorine(Compound 10C)

A mixture of compound 3C (105 mg, 0.29 mmol) and NaN3 (57 mg, 0.88 mmol)in DMF (5 mL) with Montmorillonite K-10 (50 mg) was stirred at 120° C.for 24 h. After cooling, the mixture was filtered and purified bypre-HPLC to give 10C (12 mg, 19% yield) as yellow solid. LCMS: m/z 400(M+H)+.

Example 62: Synthesis of2-(4-Methyl-[1,4]diazepan-1-yl)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid hydrazide (Compound 10D)

Compound 10D was synthesized as yellow solid (460 mg) according to theprocedure of compound 4D. LCMS: m/z 389.9 (M+H)+.

Example 63: Synthesis of2-(4-Methyl-[1,4]diazepan-1-yl)-6-(5-methyl-[1,3,4]oxadiazol-2-yl)-1,7,11b-triaza-benzo[c]fluorine(Compound 10E)

A mixture of compound 10D (45 mg, 0.11 mmol) and Triethyl orthoacetate(53 mg, 0.33 mmol) in HOAc (3 mL) was stirred at 120° C. by microwavefor 45 min. After cooling, the mixture was concentrated and purified bypre-HPLC to give compound 10E (20 mg, 41% yield) as yellow solid. LCMS:m/z 414.6 (M+H)+.

Example 64: Synthesis of2-(2-Morpholin-4-yl-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid ethyl ester (Compound 11A)

To a solution of compound SM1 (147 mg) and compound SM2 (204 mg) inethanol (10 mL) was added 2-Morpholin-4-yl-ethylamine (1 mmol) and themixture was stirred at room temperature for 20 min. The reaction mixturewas heated to reflux for 16 h. After cooling, the mixture was filteredand washed with ethanol, dried to get compound 11A as yellow solid (340mg).

Example 65: Synthesis of2-(2-Morpholin-4-yl-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid (Compound 11B)

Compound 11B was synthesized according to the procedure of compound 1A.

Example 66: Synthesis of2-(2-Morpholin-4-yl-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid hydrazide (Compound 11C)

Compound 11C was synthesized as yellow solid (40 mg) according to theprocedure of compound 4A. LCMS: m/z 406 (M+H)+.

Example 67: Synthesis of2-(2-Morpholin-4-yl-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid methylamide (Compound 11D)

Compound 11D was synthesized as yellow solid (40 mg) according to theprocedure of compound 4A. LCMS: m/z 405 (M+H)+.

Example 68: Synthesis of2-(2-Morpholin-4-yl-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid cyclopropylamide (Compound 11E)

Compound 11E was synthesized according to the procedure of compound 4A.

Example 69: Synthesis of2-(2-Morpholin-4-yl-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid methoxy-amide (Compound 11F)

Compound 11F was synthesized according to the procedure of compound 4A.

Example 70: Synthesis of2-(2-Dimethylamino-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid ethyl ester (Compound 12A)

To a solution of SM1 (1.76 g, 10 mmol) and SM2 (2.04 g, 10 mmol) in2-Methoxyethanol (100 mL) was added N1,N1-dimethylethane-1,2-diamine(2.64 g, 30 mmol) and the mixture was stirred at refluxing for 16 h. Thecrude product was purified by gel silica column (DCM/MeOH=50:1) to get1.4 g crude product. The crude product was purified by pre-HPLC to getcompound 12A as yellow solid (760 mg, 20% yield). LCMS: m/z 378 (M+H)+.

Example 71: Synthesis of2-(2-Dimethylamino-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid hydrazide (Compound 12B)

Compound 12B was synthesized as yellow solid (11 mg) according to theprocedure of compound 4D. LCMS: m/z 364.5 (M+H)+.

Example 72: Synthesis of2-(2-Dimethylamino-ethylamino)-1,7,11b-triaza-benzo[c]fluorene-6-carboxylicacid methylamide (Compound 12C)

Compound 12C was synthesized as yellow solid (20 mg) according to theprocedure of compound 4D. LCMS: m/z 363 (M+H)+.

Example 73: Synthesis of 3,7,11b-Triaza-benzo[c]fluorene-6-carboxylicacid ethyl ester (Compound 13A)

To a solution of compound SM3 (0.7 g) and compound SM2 (1.02 g) inethanol (50 mL) was added piperidine (15 mmol) and the mixture wasstirred at room temperature for 20 min. The reaction mixture was heatedto reflux for 16 h. After cooling, the mixture was filtered and washedwith ethanol, dried to get compound 13A as yellow solid (1.2 g).

Example 74: Synthesis of 3,7,11b-Triaza-benzo[c]fluorene-6-carboxylicacid (Compound 13B)

Compound 13B was synthesized as yellow solid (420 mg) according to theprocedure of compound 1A. LCMS: m/z 264.4 (M+H)+.

Example 75: Synthesis of 3,7,11b-Triaza-benzo[c]fluorene-6-carboxylicacid isopropylamide (Compound 13C)

Compound 13C was synthesized as pale yellow solid (25 mg) according tothe procedure of compound 4D. LCMS: m/z 305.5 (M+H)+.

Example 76: Synthesis of 3,7,11b-Triaza-benzo[c]fluorene-6-carboxylicacid (2-dimethylamino-ethyl)-amide (Compound 13D)

Compound 13D was synthesized as pale yellow solid (38 mg) according tothe procedure of compound 4D. LCMS: m/z 334.5 (M+H)+.

Example 77: Synthesis of 3,7,11b-Triaza-benzo[c]fluorene-6-carboxylicacid (3-methoxy-propyl)-amide (Compound 13E)

Compound 13E was synthesized as pale yellow solid (22 mg) according tothe procedure of compound 4D. LCMS: m/z 336.5 (M+H)+.

Example 78: Synthesis of 3,7,11b-Triaza-benzo[c]fluorene-6-carboxylicacid (2-morpholin-4-yl-ethyl)-amide (Compound 13F)

Compound 13F was synthesized as pale yellow solid (41 mg) according tothe procedure of compound 4D. LCMS: m/z 376.5 (M+H)+.

Example 79: Synthesis of 3,7,11b-Triaza-benzo[c]fluorene-6-carboxylicacid (2-pyrrolidin-1-yl-ethyl)-amide (Compound 13G)

Compound 13G was synthesized as pale yellow solid (31 mg) according tothe procedure of compound 4D. LCMS: m/z 360.4 (M+H)+.

Example 80: Synthesis of[1,4]Diazepan-1-yl-(3,7,11b-triaza-benzo[c]fluoren-6-yl)-methanone(Compound 13H)

Compound 13H was synthesized as yellow solid (10 mg) according to theprocedure of compound 7D. LCMS: m/z 346 (M+H)+.

Example 81: Synthesis of(4-Methyl-[1,4]diazepan-1-yl)-(3,7,11b-triaza-benzo[c]fluoren-6-yl)-methanone(Compound 13I)

Compound 13I was synthesized as yellow solid (10 mg) according to theprocedure of compound 4A. LCMS: m/z 360 (M+H)+.

Example 82: Synthesis of 3,7,11b-Triaza-benzo[c]fluorene-6-carboxylicacid (1-methyl-piperidin-4-yl)-amide (Compound 13J)

Compound 13J was synthesized as yellow solid (16 mg) according to theprocedure of compound 4A. LCMS: m/z 360 (M+H)+.

Example 83: Synthesis of(3,5-Dimethyl-piperazin-1-yl)-(3,7,11b-triaza-benzo[c]fluoren-6-yl)-methanone(Compound 13K)

Compound 13K was synthesized as yellow solid (16 mg) according to theprocedure of compound 4A. LCMS: m/z 360 (M+H)+.

Example 84: Representative Cell-Based IC₅₀ Data

Inhibitory activity on cell proliferation of representative compounds ofthe invention was determined using an Alamar Blue cell viability assayas described hereafter.

3000 cancer cells in 100 uL of cell culture media were plated in eachwell of a 96-well clear bottom, black wall cell culture-pretreatedplate.

The next day compounds are serially diluted (3-fold in cell culturemedia) across a 96-well polypropylene mother plate from row A to row F,to yield 6 concentrations (10 uM, 3.3 uM, 1.1 uM, 370 nM, 124 nM and 41nM) for each test compound. Rows G and H contain only DMSO.

Once titrations are made, the media in plates with cells were disposedand 100 μL of drug dilutions are transferred to plates with cells. Aftera ninety six-hour incubation at 37° C., 10 uL of resazurin solution fromAlamar Blue Cell Viability kit (Invitrogen, Carlsbad, Calif.) was addedto the media and cells were incubated at 37° C. for three more hours. Atthe end of this incubation the production of resofurin was measuredusing Spectrmax M2 microplate reader (Molecular Devices, Sunnyvale,Calif.)

Example 85: qRT-PCR Assay for Selective Inhibition of RNA Polymerase ITranscription

3000 cancer cells in 100 uL of cell culture media were plated in eachwell of a 96-well clear bottom, black wall cell culture-pretreatedplate.

The next day compounds are serially diluted (3-fold in cell culturemedia) across a 96-well polypropylene mother plate from row A to row F,to yield 6 concentrations (10 uM, 3.3 uM, 1.1 uM, 370 nM, 124 nM and 41nM) for each test compound. Rows G and H contain only DMSO.

Once titrations are made, the media in plates with cells were disposedand 100 μL of drug dilutions are transferred to plates with cells. Aftera ninety six-hour incubation at 37° C., 10 uL of resazurin solution fromAlamar Blue Cell Viability kit (Invitrogen, Carlsbad, Calif.) was addedto the media and cells were incubated at 37° C. for three more hours.

At the end of this incubation the production of resofurin was measuredusing Spectrmax M2 microplate reader (Molecular Devices, Sunnyvale,Calif.)

The Pol I transcription assay was used to measure the compound-dependentinhibition of the synthesis of rRNA versus mRNA. Briefly, this procedureutilizes a quantitative real time polymerase chain reaction assay(qRT-PCR) to quantify the amount of newly synthesized rRNA and mRNA incancer cells treated with the drugs. The format of this assay is thesame for all cell lines tested. Assay protocol is described hereafter.

2*10⁵ cancer cells in 2 mL of cell culture media were plated in eachwell of a 6-well clear bottom, black wall cell culture-pretreated plate.The next day compounds are serially diluted (5-fold in cell culturemedia) in 15 mL conical tubes to yield 6 concentrations (25 uM, 5 uM, 1uM, 200 nM, 40 nM and 8 nM) for each test compound.

Once titrations are made, the media in plates with cells were disposedand 2 mL of drug dilutions are transferred to plates with cells. Aftertwo-hour incubation at 37° C., the media with drug dilutions isdisposed, the cells in the plate are washed once with 2 mL of ice-coldPBS and the total RNA from cells is isolated using RNAqueous®-MicroTotal RNA Isolation Kit (Lechnologies, Carlsbad, Calif.) according tothe manufacturer's protocol) and its concentration was determined usingRibogreen reagent (Life Lechnologies, Carlsbad, Calif.).

Relative levels of 45S pre-rRNA and c-myc mRNA were measured usingApplied Biosystems' (Foster City, Calif.) proprietary primers-probe setfor c-myc mRNA and custom primers probe set (forward primer:CCGCGCTCTACCTTACCTACCT SEQ. ID 1, reverse primer:GCATGGCTTAATCTTTGAGACAAG SEQ. ID 2, probe: TTGATCCTGCCAGTAGC SEQ. ID 3)for pre-rRNA. Analysis was run on 7500HT Real Time PCR System (AppliedBiosystems, Foster City, Calif.).

Example 86: Cell-Free Pol I Transcription Assay

To measure the direct effect of representative compounds on RNAPolymerase I transcription, a nuclear extract-based assay was used.Assay protocol is described hereafter.

Compounds are serially diluted (5-fold in cell culture media) across a96-well polypropylene mother plate from row A to row E, to yield 5concentrations (50 uM, 10 uM, 2 uM, 400 nM and 80 nM) for each testcompound. Row G contained only DMSO.

Once titrations were made, the reaction mixture consisting of 30 ng/uLDNA template corresponding to (−160/+379) region on rDNA and 3 mg/mLnuclear extract isolated from HeLa S3 cells in a buffer containing 10 mMTris HCl pH 8.0, 80 mM KCl, 0.8% polyvinyl alcohol, 10 mg/mL a-amanitinwas combined with the test compounds and incubated at ambient for 20min.

Transcription was initiated by addition of rNTP mix (New EnglandBiolabs, Ipswich, Mass.) to a final concentration of 1 mM and wasincubated for one hour at 30° C. Afterwards DNase I was added and thereaction was further incubated for 2 hr at 37° C.

DNase digestion was terminated by the addition of EDTA to finalconcentration of 10 mM, followed immediately by 10 min incubation at 75°C., and then samples were transferred to 4° C. The levels of resultanttranscript were analyzed by qRT-PCR on 7500HT Real Time PCR System(Applied Biosystems, Foster City, Calif.) using the followingprimer-probe set: Pol I probe ctctggcctaccggtgacccggcta, Pol I forwardprimer gctgacacgctgtcctctggcg and Pol I reverse primerggctcaagcaggagcgcggc.

Example 87: Testing Inhibition of RNA Polymerase I and II—DrivenTranscriptions

A375 and U87-MG cells were plated in a 6-well format at 2*10^5cells/well overnight. The next day the cells were treated by a dilutionseries (6 doses total: 25 uM, 5 uM, 1 uM, 200 nM, 40 nM, 8 nM) of testcompounds. Two hours after the beginning of treatment cells were washedand lysed for RNA isolation that was performed using RNAqueous-MiniTotal RNA Isolation kit (Ambion).

The resulting RNA concentrations were determined using Quant-iTRiboGreen RNA Assay Kit (Molecular Probes). Effect on RNA Polymerase Iand RNA Polymerase II—driven transcription was assessed by monitoringresulting levels of 45S pre-rRNA and cMYC mRNA respectively. For this weperformed Taqman qRT-PCR assay using TaqMan® RNA-to-Ct™ 1-Step Kit (LifeTechnologies) with custom primer-probe set for 45S pre-rRNA (Drygin etall 2009 Cancer Res 69:7653) and Hs00153408_m1 primer-probe mix (LifeTechnologies) for cMYC mRNA. The assay was performed on AppliedBiosystems 7500 Fast Real-Time PCR System (ABI) using AbsoluteQuantitation method. The data was analyzed using GraphPad Prism(GraphPad) software.

Example 88: Pharmaceutical Activity of Representative Compounds

Representative cell proliferation inhibition from Alamar Blue assay(e.g. Example 84 herein) and Pol I and II transcription inhibition, inU-87 MG cell line, from quantitative PCR (QPCR) (Example 86 & Example87) data is provided in Table 1.

TABLE 1 Pharmaceutical activity IC50 Compound A375 A2058 Malme-3M SK-MEL28 2008 A2780 ES.2 U-87 MG PolI PolII 1A +++ 2A + 2B + 2C ++ 2D ++ 2E+++ + ++ − 2F ++ 2G +++ +++ +++ +++ +++ +++ +++ +++ +++ + 3A +++ 3B ++++++ indicates an activity of less than 1 μM; ++ indicates an activity ofgreater than 1 μM and less than 5 μM; + indicates an activity of greaterthan 5 μM and less than 10 μM; and − indicates an activity of greaterthan 10 μM.

Example 89: Pharmaceutical Activity of Representative Compounds

Representative cell proliferation inhibition from Alamar Blue assay(e.g. Example 84 herein) is provided in Table 2.

TABLE 2 Compound ES.2 Hs.294T WM-115 Hep G2 SKM-1 2G +++ +++ ++ +++ +++1A +++ +++ +++ +++ +++ 3A +++ +++ 3B +++ +++ +++ indicates an activityof less than 1 μM; and ++ indicates an activity of greater than 1 μM andless than 5 μM.

Example 90: Pharmaceutical Activity of Representative Compounds

Representative cell proliferation inhibition from Alamar Blue assay(e.g. Example 84 herein) is provided in Table 3.

TABLE 3 Compound Compound Cell line 2G 1A 3A Cell line 2G 1A 3A KG-1 ++++++ +++ DLD-1 +++ ++ +++ ML-2 +++ +++ +++ CT26 ++ + + NB4-Av +++ +++ +++LnCAP ++ + + NOMO-1 +++ +++ +++ 4T1 + + + SKM_1_AV +++ +++ +++ NCI-H1299++ ++ + B16-F10 ++ + ++ HCT-116 +++ +++ +++ AGS +++ ++ +++ A549 ++ + +MCF7 ++ ++ + PC-3 +++ ++ ++ MDA-MB-231 +++ ++ +++ HT-1080 ++ ++ + +++indicates an activity of less than 1 μM; ++ indicates an activity ofgreater than 1 μM and less than 5 μM; + indicates an activity of greaterthan 5 mm and less than 10 μM.

Example 91: Pharmaceutical Activity of Representative Compounds

Representative cell proliferation inhibition from Alamar Blue assay(e.g. Example 84 herein) is provided in Table 4.

TABLE 4 Pharmaceutical activity in A375 cell line Compound IC50 CompoundIC50 Compound IC50 Compound IC50 Compound IC50 Compound IC50  4F *** 11D** 13D *** 3F *** 12C ***  7C —  4C ***  4E ** 13F * 3G **  5B *  7F **10E ***  7D — 13G *** 3H ***  5C **  7G —  9B **  8B ***  3C *** 3I ** 5E **  7I ** 12B **  8D ***  3D *** 4A *  5D * 13E * 11C * 13C *  3E **4D ***  6A *  7B — *** indicates an IC50 of less than 1 μM; ** indicatean IC50 of greater than 1 μM and less than 5 μM; * indictes an IC50greater than 5 μM and less than 10 μM; — indicates an IC50 of greaterthan 10 μM

Example 92: Pharmaceutical Activity of Representative Compounds

Representative cell proliferation inhibition from Alamar Blue assay(e.g. Example 84 herein) is provided in Table 5.

TABLE 5 IC₅₀ of compound 2G in ovarian cell lines kuramochi JHOC7 JHOC9JHOM1 EF-21 TOV112D *** *** *** *** *** *** *** indicates an IC₅₀ ofless than 1 μM

Example 93: Pharmaceutical Activity of Representative Compounds

Representative Pol I and II transcription inhibition, in A375 MalignantMelanoma cell line, from quantitative PCR (QPCR) (Example 85 and Example87) data is provided in Table 6.

TABLE 6 Pol I and Pol II IC₅₀ Compound Pol I Pol II  3A **** ND  3D ****** 12C *** ** 13D **** ****  8B ** * 10E ** **** **** indicates an IC₅₀of less than 0.5 μM; *** indicates an IC₅₀ between 0.5 μM and 1 μM; **indicates an IC₅₀ of greater than 1 μM; ND means not determined

The above description of the disclosed embodiments is provided to enableany person skilled in the art to make or use the invention. Variousmodifications to these embodiments will be readily apparent to thoseskilled in the art, and the generic principles described herein can beapplied to other embodiments without departing from the spirit or scopeof the invention. Thus, it is to be understood that the description anddrawings presented herein represent a presently preferred embodiment ofthe invention and are therefore representative of the subject matterwhich is broadly contemplated by the present invention. It is furtherunderstood that the scope of the present invention fully encompassesother embodiments that may become obvious to those skilled in the artand that the scope of the present invention is accordingly not limited.

What is claimed is:
 1. A method for treating cancer in a subject,comprising the administration of an effective amount of a compoundhaving the structure of Formula II(A), II(B), II(C), or II(D), or apharmaceutically acceptable salt, hydrate or tautomer thereof, whereinthe cancer is of the breast, lung, colorectum, liver, pancreas, lymphnode, colon, prostate, brain, head and neck, skin, kidney, blood orheart; wherein the compound of Formula II(A), II(B), II(C), or II(D) hasthe following structure:

wherein: Y is an optionally substituted 5-6 membered carbocyclic orheterocyclic ring; X₁ is an optionally substituted C₁-C₈ alkyl, C₂-C₈heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group,optionally substituted with one or more halogens, ═O, CF₃, CN, OR₇,NR₈R₉, SR₇, SO₂NR₈R₉, C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl,C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl,C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, orC₆-C₁₂ heteroarylalkyl group; wherein each R₇, R₈ and R₉ isindependently selected from H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, ═O, ═N—CN,═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂,NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and NO₂; whereineach R′ is independently H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl,C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, orC₆-C₁₂ heteroarylalkyl, each of which is optionally substituted with oneor more groups selected from halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O; wherein two R′ can belinked to form a 3-7 membered ring optionally containing up to threeheteroatoms selected from N, O and S; or X₁ is H, NR₂R₃, SOR₂, SO₂R₂,SO₂NR₂R₃, NR₂SO₂R₃, NR₂CONR₂R₃, NR₂COOR₃, NR₂COR₃, CN, COOR₂, COOH,CONR₂R₃, OOCR₂, COR₂, or NO₂, wherein each R₂ and R₃ is independentlyselected from H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂heteroarylalkyl, each of which is optionally substituted with one ormore groups selected from halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O; wherein R₂ and R₃ groupson the same atom or on adjacent atoms can be linked to form a 3-8membered ring, optionally containing one or more N, O or S atoms; andeach R₂ and R₃ groups, and each ring formed by linking R₂ and R₃ groupstogether, is optionally substituted with one or more substituentsselected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′,SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂,OOCR′, COR′, and NO₂; wherein each R′ is independently H, C₁-C₆ alkyl,C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of whichis optionally substituted with one or more groups selected from halo,C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy,amino, and ═O; wherein two R′ can be linked to form a 3-7 membered ringoptionally containing up to three heteroatoms selected from N, O and S;X₂ is H, halogen, CF₃, CN, OR₇, NR₈R₉, SR₇, SO₂NR₈R₉; C₁-C₁₀ alkyl,C₁-C₁₀ heteroalkyl, C₂-C₁₀ alkenyl, or C₂-C₁₀ heteroalkenyl, each ofwhich is optionally substituted with one or more halogens, ═O, anoptionally substituted 3-7 membered carbocyclic or heterocyclic ring,wherein each R₇, R₈ and R₉ is independently selected from H, C₁-C₆alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl,C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each ofwhich is optionally substituted with one or more groups selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′, SO₂NR′₂,NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′,COR′, and NO₂; wherein each R′ is independently H, C₁-C₆ alkyl, C₂-C₆heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl, each of whichis optionally substituted with one or more groups selected from halo,C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl, hydroxy,amino, and ═O; wherein two R′ can be linked to form a 3-7 membered ringoptionally containing up to three heteroatoms selected from N, O and S,and wherein R₇ is not hydrogen when X₂ is OR₇; Z₂, Z₃ and Z₄ are eachindependently CH or CR₁; wherein each R₁ is independently an optionallysubstituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl, C₂-C₈heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl, C₂-C₈heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂heteroarylalkyl group; or each R₁ is independently H, halo, CF₃, OR₂,NR₂R₃, NR₂OR₃, NR₂NR₂R₃, SR₂, SOR₂, SO₂R₂, SO₂NR₂R₃, NR₂SO₂R₃,NR₂CONR₂R₃, NR₂COOR₃, NR₂COR₃, CN, COOR₂, COOH, CONR₂R₃, OOCR₂, COR₂, orNO₂; or each R₁ is independently —W, -L-W, —X-L-A; wherein X is NR₆, O,or S; wherein R₆ is H, optionally substituted C₁-C₈ alkyl, C₂-C₈heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group; whereinR₆ can be linked to R₄ or R₅ to form a 3-8 membered ring optionallysubstituted with one or more substituents selected from halo, ═O, ═N—CN,═N—OR′, ═NR′, OR′, N(R′)₂, SR′, SO₂R′ SO₂NR′₂, NR′SO₂R′, NR′CONR′₂,NR′COOR′, NR′COR′, CN, COOR′, CON(R′)₂, OOCR′, COR′, and NO₂;  whereineach R′ is independently H, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl,C₂-C₆ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, orC₆-C₁₂ heteroarylalkyl, each of which is optionally substituted with oneor more groups selected from halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆acyl, C₁-C₆ heteroacyl, hydroxy, amino, and ═O;  wherein two R′ can belinked to form a 3-7 membered ring optionally containing up to threeheteroatoms selected from N, O and S; each W is independently anoptionally substituted 4-7 membered azacyclic ring, optionallycontaining an additional heteroatom selected from N, O and S as a ringmember; L is a bond, C₁-C₁₀ alkylene, C₁-C₁₀ heteroalkylene, C₂-C₁₀alkenylene or C₂-C₁₀ heteroalkenylene linker, each of which isoptionally substituted with one or more substituents selected from thegroup consisting of halogen, oxo (═O), or C₁-C₆ alkyl; and A isheterocycloalkyl, heteroaryl or NR₄R₅, wherein R₄ and R₅ are eachindependently H, optionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl,C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl,C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl group; wherein R₄ and R₅ areoptionally linked to form a 3-8 membered ring, optionally containing oneor more N, O or S; and each R₄ and R₅ groups, and each ring formed bylinking R₄ and R₅ groups together, is optionally substituted with one ormore substituents selected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′,N(R′)₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN,COOR′, CON(R′)₂, OOCR′, COR′, and NO₂;  wherein each R′ is independentlyH, C₁-C₆ alkyl, C₂-C₆ heteroalkyl, C₁-C₆ acyl, C₂-C₆ heteroacyl, C₆-C₁₀aryl, C₅-C₁₀ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl,each of which is optionally substituted with one or more groups selectedfrom halo, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₁-C₆ acyl, C₁-C₆ heteroacyl,hydroxy, amino, and ═O;  wherein two R′ can be linked to form a 3-7membered ring optionally containing up to three heteroatoms selectedfrom N, O and S.
 2. The method of claim 1, wherein X₁ is CN, COOR₂, orCONR₂R₃ and X is NR₆.
 3. The method of claim 1, wherein each R₁ isindependently an optionally substituted C₁-C₈ alkyl, C₂-C₈ heteroalkyl,C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈ alkynyl, C₂-C₈ heteroalkynyl,C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl, C₅-C₁₂ heteroaryl, C₇-C₁₂arylalkyl, or C₆-C₁₂ heteroarylalkyl group, or each R₁ is independentlyhalo, CF₃, OR₂, NR₂R₃, NR₂OR₃, NR₂NR₂R₃, SR₂, SOR₂, SO₂R₂, SO₂NR₂R₃,NR₂SO₂R₃, NR₂CONR₂R₃, NR₂COOR₃, NR₂COR₃, CN, COOR₂, COOH, CONR₂R₃,OOCR₂, COR₂, or NO₂.
 4. The method of claim 2, wherein each R₁ isindependently —W, -L-W, —X-L-A; wherein X is NR₆, O, or S; W is anoptionally substituted 4-7 membered azacyclic ring optionally containingan additional heteroatom selected from N, O and S as a ring member; L isa bond, C₁-C₁₀ alkylene, C₁-C₁₀ heteroalkylene, C₂-C₁₀ alkenylene orC₂-C₁₀ heteroalkenylene linker, each of which is optionally substitutedwith one or more substituents selected from the group consisting ofhalogen, oxo (═O), or C₁-C₆ alkyl; and A is heterocycloalkyl,heteroaryl, or NR₄R₅; wherein R₄ and R₅ are independently H, C₁-C₈alkyl, C₂-C₈ heteroalkyl, C₂-C₈ alkenyl, C₂-C₈ heteroalkenyl, C₂-C₈alkynyl, C₂-C₈ heteroalkynyl, C₁-C₈ acyl, C₂-C₈ heteroacyl, C₆-C₁₀ aryl,C₅-C₁₂ heteroaryl, C₇-C₁₂ arylalkyl, or C₆-C₁₂ heteroarylalkyl group;wherein R₆ is H or C₁-C₈ alkyl; or wherein R₆ is linked to R₄ or R₅ toform a 3-8 membered ring optionally substituted with one or moresubstituents selected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, N(R′)₂,SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′COOR′, NR′COR′, CN, COOR′,CON(R′)₂, OOCR′, COR′, and NO₂.
 5. The method of claim 1, wherein R₆ isH or C₁-C₈ alkyl; or R₆ is linked to R₄ or R₅ to form a 3-8 memberedring; and Y is an optionally substituted 5-6 membered carbocyclic or 5-6membered heterocyclic ring.
 6. The method of claim 1, wherein: X₁ is CN,COOR₂, or CONR₂R₃.
 7. The method of claim 6, wherein: X₂ is H, halogen,CF₃, CN, OR₇, NR₈R₉, or C₁-C₁₀ alkyl.